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REGN 7257

The creation of new drugs has been greatly improved with new discoveries in biology, chemistry, and the human body. However, the newfound complexity of synthesizing and marketing these drugs for medical applications demands a more thorough understanding of the specific interactions between molecules as well as the legal framework for monetization. These hurdles can be seen with Regeneron's REGN 7257, an experimental drug for the treatment of aplastic anemia. A substantial amount of research went into the drug before trials even began. While the drug is already in phase I/II of its clinical trials, it still has a long path ahead of it before the public has access to it.

The REGN 7257 drug, while still in development, is relatively easy to synthesize once all the components are together. The two main components of the REGN 7257 drug is an antibody or an antigen-binding fragment and the interleukin-2 gamma chain which bind to each other at the epitope. Once extracted the interluekin-2 gamma chain and the antibody are then dissolved in a solution filled with water and some buffer ions and allowed to mix at a moderate temperature anywhere in the range of 25^oC and 40^oC. Different temperatures will give the antibody different dissociation constants which will reflect how well the antibody will actually bind to the gamma chain. Once the binding has happened at the epitope the drug has been synthesized.

Currently, the REGN 7257 drug has a pending patent under evaluation that was filed as of 2021. As it is undergoing this process, the patent examiners are working with representatives of the REGN 7257 company to negotiate the novelty and the proprietary claims on the legal document. Under this process, the REGN-7257 work will be measured against prior art in the field in order to determine the validity of the patent for this material. Since REGN-7257 is a newly innovated drug, it does benefit from having the U.S. safe harbor expectation that allows it to undergo clinical trial to prove that the product works while still maintaining protection for the proprietary material it will inevitably contribute. Like most other medications, this material is best protected using a patent that explicitly lists the infrangible claims which will eventually allow the owners of the patent to market their material and own the exclusive rights to create and sell the medication they are currently testing.

In order to test REGN 7257, clinical trials including only patients with aplastic anemia must be conducted. The diagnostic process of aplastic anemia is fairly uncomplicated. Symptoms that may lead to further testing for aplastic anemia include frequent bruising, nosebleeds, fatigue, or abnormal heart rate. Further testing on a patient exhibiting these symptoms may include running blood tests such as a Complete Blood Count or basal metabolic panel. A bone marrow biopsy may also be used in diagnosis of aplastic anemia. With the REGN-7257 drug, the main focus is treatment of immunocompromised patients, or patients with an autoimmune disorder. Aplastic anemia is sometimes caused by such a disorder, but in other patients without an immune component, treatments such as blood transfusions or Bone Marrow Transplant can be used in hopes of treating or even curing the disease. These treatments are either non-applicable or unsustainable for patients having aplastic anemia with an immune component, therefore other options are being explored for these patients, with the prime example being the creation and testing of the drug REGN 7257.

In order for REGN-7257 to be properly tested, it requires that the patients are confirmed to be diagnosed with Aplastic Anemia. This confirmation of Aplastic Anemia is crucial as Aplastic Anemia can be mistaken for other disorders such as Paroxysmal Nocturnal Hemoglobinuria (PNH). The confirmation of the diagnosis of Aplastic Anemia can be done via many tests; however, the most common are a complete blood count (CBC) and/or a bone biopsy exam. These two options can provide a comprehensive diagnosis for Aplastic Anemia with only minor procedures such as the bone marrow extraction procedure.

As REGN 7257 is a chemical that works by binding to T-cells, it follows a path that starts by initially bindings on the inner surface of the cell membrane as opposed to an outer enzyme. These T-cell proteins are known for stemming from the bone marrow and working alongside macrophages in the body to activate the t-cell immunity in target organs. They are focused in their role in fighting against infections and diseases in the body, but people that struggle with autoimmune diseases such as aplastic anemia lack that ability to undergo constant hematopoiesis of white blood cells, and the REGN 7257 medicine is aimed at fixing just that.

REGN 7257 hopes to mitigate the symptoms of aplastic anemia by suppressing the patient's immune system. This is accomplished by specifically targeting immune cells using antibody-mediated drug delivery. This form of drug delivery harnesses monoclonal antibodies and receptors naturally found in the body to ensure REGN 7257 arrives to B and T lymphocytes. These cells are the primary carriers of the cytokine receptor interleukin-2. The cargo carried by the drug then suppresses the receptors by binding to them. By blocking any other interaction with the IL-2 receptors, the suppressed immune cells can no longer activate similar cells or spur their growth.

The path REGN 7257 has taken initially appears daunting, but familiarity with scientific jargon and technical experience can make the process easier to embark on. To help our readers, attached to this Wikipedia article is a list of commonly used terms and their definition as well as some professional communities that emphasize the drug making process.

Synthesis

Interleukin-2 Gamma and Antibodies

This is a 3D representation of the Interleukin-2 protein

The Interleukin-2 (IL-2) protein is a T cell-derived cytokine and it targets numerous cells to induce growth and functional activation. In humans, the IL-2 receptor consists of 3 subunits the alpha, beta, and gamma chains. The alpha and beta chain of IL-2 are more common amongst other proteins so scientists have been able to figure out their functionality in the IL-2 protein. However, the gamma chain has been researched to a lesser extent, but it has still been found to increase binding affinity for the IL-2 protein. Mutations of the gamma chain have also been seen to cause X-linked severe combined immunodeficiency (XSCID). This being said, since there is a gap of knowledge of the gamma chain a lot of experimental drugs have been developed using it to better understand its properties. For the synthesis of the REGN 7257 drug the two main components are the IL-2 gamma chain and some antibodies or antigen binding fragments. These antibodies used in the synthesis of the drug can be any antibodies in the body assuming they are from a human.

Protein-Ligand Interactions

For the actual synthesis of the REGN 7257 drug it is relatively simple once all of the constituents are received. The synthesis begins with preparing a solvent consisting of water and some buffer ions and then the solute of the system will be the protein (IL-2 gamma chain) and the ligand molecules (antibodies or antigen binding fragments). This solute-solvent system can then be heated up very slightly anywhere between 25^oC and 37^oC to ensure that the dissociation constant remains low so that the antibodies can bind to the IL-2 gamma chain. The lower the dissociation constant the higher the affinity for binding will be between the antibodies and the gamma chain. In the range of 25-37^oC the dissociation constant ranges from 2.75 x 10^-9M and 3.53 x 10^-7M which is an extremely low dissociation constant. The antibodies present in the solution will bind specifically to the epitope of the IL-2 gamma chain which basically means there is a specific point on the gamma chain where the antibodies can attach to due to their geometry. Once the epitope of the gamma chain has been found there are three general ways in which the antibody can bind to it: the lock-and-key method, the induced fit method, and the conformation selection method. The lock-and-key method assumes that there is a specific shape in the gamma chain where the antibody fits perfectly which is very unlikely. The induced fit method assumes that the epitope on the gamma chain is flexible and can accommodate any variable difference in the shape of the antibody. Finally, the conformational selection method assumes that the epitope on the gamma chain is not just one shape but, in fact, it changes over time and the antibody should bind bind at the correct shape. Epitope mapping can be used to determine where and how the antibodies will bind to the gamma chain but overall, the synthesis of the drug is not very complicated.

3D results of epitope mapping

Epitope Mapping

Epitope mapping is an essential part of antibody development and for understanding how antibodies work during the binding process. Although not required to understand for the synthesis epitope mapping can be very useful for researchers to understand where antibodies bind to because differences in binding can lead to different effects on the body. Epitope mapping can also be used to select specific antibodies that target specific points of a protein so that the desired effects can come through. Epitope mapping won't only tell you where an epitope is but what type it is as well. An epitope can be linear, conformation, or discontinuous and all the different types of epitopes can have different properties and allow for different antibodies to connect to them. The IL-2 gamma chain, for example, consists of mainly discontinuous epitopes which allow for a specific type of antibody. Epitope mapping is very cost-effective but the major problem with it is that it take a while to fully track all the epitopes of a protein. Since the system has to scan every sequence of amino acids depending on the protein size epitope mapping can take anywhere from 5-12 weeks to complete.

Legal protection

United States Patent and Trademark Office

World Trade Organization Members: Where Intellectual Property Law is practiced at the same standard

Protecting Intellectual Property

There are many ways a company can choose to protect their intellectual property, and globally speaking, the means of protecting Intellectual property in the United States is close to uniform internationally. Generally speaking the categories include patents, trademarks, trade secrets , and copyright. Under the case of REGN-7257, the most commonly practiced means of protecting its material would likely be a patent. According to the Yale Law Journal, the reasoning behind this would be that trade secret law, though more severe for misappropriation, might be unrealistic to maintain. Patents offer disclosure and protection which allows for the researchers to disclose their material and still be able to obtain exclusive rights, while a trade secret, the other valid option, would require that the researchers themselves protect their own data under security measures within their own company. Not only is this inherently difficult to do, but the novel drug is undergoing clinical trials and will eventually have to petition for FDA approval, and with these factors in mind it is unreasonable to maintain the chemical formula and process a trade secret since there is inevitable disclosure. Therefore, at this step, it would be best for the legal counsel to file a probationary patent pending clinical trial results so they can have the freedom to disclose the material but also the safety of the patent.

Example patent with claims it protects

Safe harbor for drug manufacturing

The REGN-7257 drug is currently undergoing a clinical trial to test its efficacy rate for people with aplastic anemia. During this time, the company must also begin the filing process to protect the invention they are working on. According Oxford University Press Law and Intellectual property journal, due to the nature of the industry, there has been a new safe harbor allocation in the United States to account for disclosure during a clinical trial for new medications and how that would affect publication and market approval. REGN-7257 is a perfect example of how this safe harbor would be beneficial to the inventing party. One of the basic qualifications to be issued a patent is for the party to prove it works and is going to contribute to society. This causes an issue with drug manufacturing since there is no feasible way to ensure it will be effective without trials, and no way to perform trials without disclosing what material will be tested. This safe measure provides space for the inventors to ensure their claims will work, but also protect the information at the same time.

USPTO Patent Search Platform

All patents in the US must be processed by the United States Patent and Trademark office. As the official and exclusive handlers of this, the USPTO provided a database that allows for people to search for pending patents and current patents. The reason this resource is useful is because of the first to file ruling that dictates that an inventor's right to a patent is issued by the party that is the first to the filing office. Additionally, previous patents can be used as prior art artifacts which could negate future patents as being obvious or anticipated based on what has already been published and patented. Additionally, a patent’s claims are the entire basis of being able to exercise the right to sue on infringement and because of the quid pro quo basis of patent. In exchange for exclusive rights, the invention must be disclosed to the public domain which can be accessed through this USPTO platform.

USPTO Patent Application

This platform is used for inventors to submit their application. The first party to file their patent application will be given that filing date to litigate any future infringement disputes. The patent application itself is composed of several different areas that could expedite the process to issuance. There are priority and accelerated options which cost more, but allow for the patent to be examined and issued quicker. Though the validity is from filing date the patent cannot be used to file an injunction or infringement suite until it has been completely issued. This is why expedited processes are valuable for those in a lucrative innovative field. This platform provided by the USPTO is accessible meaning if the inventor does not already work with legal counsel, it is still possible to apply for the patent. Generally speaking, these fees are $8,000-$10,000 for the application itself, but it is recommended to allocate up to $15,000 to account for legal fees during the negotiation process.

Diagnosis

Aplastic Anemia - Overview

Diagram of Central Venous Catheter - often used for patients needing frequent blood or medicine transfusions

Aplastic anemia occurs when the body's bone marrow fails to produce more red blood cells. Generally, the condition can be caused by many different sources, but the most severe form of aplastic anemia comes from the body's immune system attacking it's own bone marrow. Aplastic anemia can range from moderate to severe, but often afflicted patients must receive frequent blood transfusion, if not a more invasive treatment. Other treatment options include bone marrow transplant, medical therapy options including immune system suppression, or other clinical options such as the REGN 7257 which aim to provide less-invasive alternative treatments.

Symptoms of aplastic anemia that may lead to further diagnostic testing include fatigue, nosebleeds and bleeding gums, frequent infections, repeated and easy bruising, and several other less severe symptoms.

There are several causes of aplastic anemia, the most severe form coming from a type of autoimmune condition, in which the immune system attacks the bone marrow of the body. Other causes include exposure to certain chemicals or medications -- which can range from accidental toxic chemical exposure to ingested medications, to chemotherapy and radiation treatments. Aplastic anemia can also be a side effect of pregnancy, or a viral infection that attacks the bone marrow or blood contents. In some cases, aplastic anemia can be related to rare disorders such as paroxysmal nocturnal hemoglobinuria- a condition in which the red blood cells break down too quickly, or a disorder called Fanconi's Anemia.

Diagnostic and Treatment Options

Often, aplastic anemia is diagnosed via blood testing, such as a Complete Blood Count (CBC) or more complicated procedures such as Bone Marrow Biopsy. Treatment options are much the same, most procedures surrounding blood infusions, bone marrow transplants, and medications or treatments affecting the blood and blood cells.

Process of Allogenic Bone Marrow Transplant

One of the more extreme treatments for aplastic anemia includes Bone Marrow Transplant. In patients with severe aplastic anemia, bone marrow transplant is often one of the primary treatment options. In this transplant, healthy stem cells can be donated from you own body (autologous transplant), or healthy stem cells from a matching donor can be used (allogenic transplant). In this transplant, conditioning will need to be done on current, unhealthy bone marrow cells, which generally includes chemotherapy and radiation treatments. After the patient is prepared, stem cells from donor marrow are either injected into a vein connected to the marrow itself, or diffused into a central line such as the Central Venous Catheter (seen above). After injected with the donor marrow, donor cells begin to join the original marrow and being producing healthy blood cells.

In a study done on patients with aplastic anemia who received allogenic bone marrow transplants from siblings with matching Human Leukocyte Antigens (HLA), a 10-year survival rate of 64% was found. A much higher survival rate (up to 90% of patients with severe aplastic anemia cured) can be achieved by minimizing immunosuppressive therapy before transplant, and using specific chemotherapy and radiation treatments in patient conditioning.

Sources used in diagnosis

In the medical field, many treatments, studies, and diagnosis are a collaborative effort between multiple doctors and physicians. The internet has only expanded on the resources and collaborations between medical professionals, and sources such as PubMed and Upto Date can aid in medical diagnoses and sharing of information.

Logo for PubMed

PubMed is a website run by the National Health Institute, giving doctors, researchers, and patients the ability to investigate everything from medical condition information to knew medical breakthroughs. Through PubMed, doctors are able to share newfound knowledge that can be used in hospitals world-wide, regardless of the original doctor or researcher being present to aid with teaching new knowledge. PubMed includes access to full-text articles, informational reviews, new research, and even information on clinical studies open to patients who qualify. Using PubMed gives doctors an avenue to brush up on less-familiar procedures, conditions, and even make medical discoveries from the information available. Information found on PubMed is generally related to biomedical and life sciences literature, and can not only be used by practicing physicians, but also by students and researchers aiming to learn more about a specific topic or field. PubMed acts as a library full of medical and biological information, and links a vast source of knowledge to millions of doctors, researchers, students, and patients worldwide.

Upto Date is a subscription-based medical site that acts as a support for practicing physicians. The software offers several resources, from medical information from trials, drug interaction information, patient records, and even medical calculators. The software originally included only information concerning the nephrology specialty (cases concerning health of the kidneys), but now includes information and resources for over twenty other medical specialties. Information, records, and advice found on Upto Date is written by over 7,100 physicians, and all work is both edited and peer reviewed before being published on the site. Upto Date includes resources not only for medical diagnoses and patient symptoms, but includes resources for furthering medical education, including the ability to earn additional medical certifications. Upto Date also offers resources and advice on starting medical businesses, such as private practices, management strategies, and much more. Upto Date acts as a multi-service dictionary and school for physicians around the world, and is utilized in multiple countries and hospitals as a primary resource for information and problem solving.

Diagnosis Mechanisms

There are two main ways in which a patient could be tested for Aplastic Anemia: complete blood count, and bone marrow biopsies. The main purposes of these tests is to confirm a diagnosis of aplastic anemia, look for its cause, find out how severe it is, rule out other conditions that may cause similar symptoms, and check for paroxysmal nocturnal hemoglobinuria (PNH).

Complete Blood Count (CBC)

Formed Elements within the Blood

A complete blood count, or CBC, is usually the first type of test conducted when a patient is suspected of having Aplastic Anemia; however, it can detect a variety of disorders with the blood. A complete blood count tests for six major components within the blood: red blood cells, white blood cells, hemoglobin, hematocrit, and platelets. CBCs are fairly easy to administer with only a blood draw being required from the patient. Complete Blood Counts are measured in either cells per Liter (cell/L) or grams per deciliter (g/dL). To diagnose Aplastic Anemia, physicians often look at the hematocrit and hemoglobin results of a CBC, low hematocrit and/or hemoglobin levels could indicate anemia

Normal Ranges for a CBC

Test	Normal Range Results
Red Blood Cell	Adult Men: 5 to 6 million cells/mcL Adult Women: 4 to 5 million cells/mcL
White Blood Cell	4,500 to 10,000 cells/mcL
Platelets	140,000 to 450,000 cells/mcL
Hemoglobin (varies with altitude)	Adult Men: 14 to 17 gm/dL Adult Women:12 to 15 gm/dL
Hematocrit (varies with altitude)	Adult Men: 41% to 50 % Adult Women: 36% to 44%
Mean corpuscular volume	0 to 95 femtoliter

Bone Marrow Biopsy

Sites for Potential Bone Marrow Biopsy Injection

Due to the Aplastic Anemia being a condition affecting the bone marrow, analyzing the marrow itself can be a good source of diagnosis for Aplastic Anemia. Bone marrow aspiration and biopsies are two different procedures; however, when combined can be a great characterization of the health and status of a patient's bone marrow health. After the bone marrow extraction procedure, tests are run on the bone marrow sample in order to determine whether the patient has Aplastic Anemia, these tests include looking for a lack of red blood cells, or a lack of iron.

Bodily Response to REGN 7257

How Drugs Act on the Body

An example of a protein molecule that can be considered a binding site for chemicals.

A drug is any chemical introduced into the body that makes a physiological change in function. So, when the REGN7257 enters the body it does not create new functions or metabolic pathways but instead alters existing cascades of activity. The drug will then attach to an enzyme or receptor of some kind and act as an inhibitor essentially causing it to prevent “access from its normal substrate material,” (Georgina 2011). In the case of a chemical binding to a receptor however, the process is different in which the drug binds to the designated receptor on the inner surface of the cell membrane and activates changes in activity from there. REGN7257 is a chemical mediator that binds to T-cell receptors, so it follows the methods of the second format, and changes in the body occur from changes in T-cell activity cascades. T-cells in their normal function are known for their role in the immune system and help fight infections and diseases as they stem from the bone marrow, so an alteration of this activity with medicine would increase that rate ideally for patients that struggle with autoimmune diseases caused by less than optimal T-cell function.

T-cell Mediated Immunity

T-cells will leave the thymus and enter the bloodstream to search for a binding antigen. In our case it would be the injected REGN 7257 which attaches instead of a human antibody, and once the cell is induced function to provide lines of defense in the immune system. A group of these T-cells also further specialize and differentiate into “cytotoxic t-cells that kill infected target cells” (Janeway et al. 2001) by carrying them to the cell surface and neutralize them. This is what is meant by a primary immune response and what the REGN 7257 chemical wishes to perform when bound to the T-cell in individuals with autoimmune diseases such as aplastic anemia. While cytotoxic t-cells are focused on killing infected bacteria and cells, the body follows up by another differentiated T-cell group called armed effector T cells that provide immune memory to the body allowing it to protect itself quicker from the same pathogen harming it. Individuals that have the condition of aplastic anemia now longer have a functioning bone marrow to create new cells that do this job and therefore are prone to so many diseases and once again the chemical injected through the REGN 7257 treatment focuses on aiding limited T-cells accomplish that job.

Blood samples can be grouped and separately analyzed for protein samples such as globulin

Globulin Test

Globulins are protein groups in the blood made by the immune system and sent to the liver. They function to protect against infection while also maintaining normal function in the liver. Testing for globulins in the blood can measure the levels present and a low sing can be a noticeable indicator for liver disease and damage to the immune system due to these proteins not being created at an optimal rate. The test can provide information as to how well the immune system is working by protein levels that are created and inefficiencies can correlate to disorders of the immune system such as aplastic anemia. While low globulin levels can indicate liver disorders and immune system failure, too high of a level can indicate similar issues such as infection or immune disorders which would cause the body to essentially try and produce as many infection fighting proteins as it can in the body.

Therapeutic Drug Monitoring

An example of how the white blood cells stand as a line of defense against anything foreign

The body naturally focuses on attacking foreign substances so there could be a chance that some individuals receiving the REGN 7257 drug do not adjust well to it by their body trying to fight it to exit the system. Even though these are individuals with autoimmune disorders there are still a number of T-cells willing to act as a line of defense from the ‘invading’ substance. In this case immunosuppressive therapy can be used as a way to suppress the immune system so the drug can attach to the T-cell receptors and attempt to work. While it seems slightly counterintuitive to decrease the immune system of an individual with a low immune system already, it is almost like making it worse in an attempt to make it much better. Drug monitoring not only allows proper concentrations of the immunosuppressant to be administered but it also stays on the look out of what concentration of the chemical needs to be injected as well and keeps the body in a balance of both.

Antibody-Mediated IL-2 Immunosuppressive

Antibody-Mediated Drug Delivery

An example of the specific nature of antibodies and receptors

Conventional drug delivery methods are plagued by problems such as non-specific interactions, off-site toxicity, and a long period of time and challenges in the development of new drugs molecules. Thus, a new approach to deliver has become popular and is the subject of many new drugs currently undergoing clinical trials. Targeted drug delivery harnesses the specific targeting and interactions between antibodies and antigens/receptors to engineer drugs that interact only with their intended target. First mentioned by Dr. Paul Ehrlich, the antibody was envisioned as a “Magic Bullet” fired at a specific antigen/receptor as to not harm any surrounding healthy cells and tissue.

REGN 7257 uses the specific targeting of the anti-interleukin 2 antibody. The IL 2 cytokine is found on activated B and T cells. Thus, the anti-IL 2 antibody will direct the immunosuppressant drug specifically to the cells causing the most harm to patients with aplastic anemia. Because the antibody is specifically designed to target the IL 2 cytokine, no other cells and tissues can bind to the drug delivery system allowing then to avoid the adverse effect of the drug.

Interleukin-2

Initially described as an immunostimulatory factor, interleukin-2 (IL-2) receptor subunit has shown sophistication needed in lymphocyte development, homeostasis, and immunosuppressive functions. The existence of IL-2 was first supported in 1976 with the receptor being discovered in the 1980s, thus completing the first type 1 cytokine/receptor complex. Prolonged T cell cultures revealed IL-2’s role in facilitating T cell growth and reversal of anergy (a state in which the immune system is unable to mount a normal immune response against a specific antigen). Further tests in the 1990s showed a relationship between the lack of IL-2 and lymphadenopathy (the uncontrolled proliferation of peripheral activated T cells) and other signs of immunodeficiency. These tests established the immunosuppressive mechanisms of IL-2 which are exploited by REGN 7257 for treating aplastic anemia.

IL-2 has a sensitive balance between regulating both effector and regulatory T lymphocytes. Thus, fine-tuning the functionally contrasted T cell subsets is the primary goal of IL-2 based immunotherapies. Specifically, the immunosuppressive nature of REGN 7257 aims at selectively expanding Tregs (a specialized subpopulation of T cells that act to suppress immune response, thereby maintaining homeostasis and self-tolerance) over effector T cells. This is accomplished by the Tregs constitutively displaying the IL-2Rαβγ trimer (a macromolecular complex formed by three, usually non-covalently bound, macromolecules like proteins or nucleic acids).

Gel Electrophoresis

An image of a DNA gel electrophoresis

Gel electrophoresis is a powerful tool and was involved in the devolvement of REGN 7257. Gel electrophoresis is a technique commonly used in labs to isolate specific molecules, like proteins, according to their size. Separation is accomplished by running a current through an agarose gel. The agarose gel is prepared using varying concentrations of agarose powder depending on the size of the molecule. Because the gel is a permeable matrix, the higher concentrations create tighter and tighter holes in which the molecules can travel through. The molten gel is then placed into a casting tray with a comb inserted at one end to make wells for the samples to be pipetted into. Once the gel has solidified, the comb can be removed, and the samples inserted. The samples are prepared by adding a dye beforehand to prevent the sample from escaping the wells as well as to allow the migration of the sample to be observed through the gel. Finally, a protein ladder is loaded into the gel. The protein ladder has fragments of known lengths to help approximate the size of the collected sample. Passing a current through the gel then pulls all the prepared samples down the gel. Once samples have been separated, the specific molecules can be removed and purified from the gel.

Monoclonal Antibody Production

A visual of the monoclonal antibody production

To produce the monoclonal antibody (mAb) used in REGN 7257, the traditional production process begins by generating mAb-producing cells. These hybridomas are created by fusing together myeloma cells, immortalized cells with unique unlimited growth property, with B lymphocytes that express the desires antibody. These splenocytes are commonly supplied by mice immunized specifically for an antibody. Their spleen is then isolated and prepared for in vitro hybridoma production. Once the B lymphocytes and myeloma cells are prepared, they are fused together with the aid of polyethylene glycol. After fusion, the resulting clones are screened and selected based on their antigen specificity and immunoglobin class. Further tests are carried out to validate and characterize the antibody of these colonies. One test, the ELISA, tests the antibody by using specialized enzymes that attach to known antibodies. When all tests are finished, the clone production of the antibody is scaled up to meet the desired amount. Through this process, monoclonal antibodies can be produced on a commercial scale.

Lists of Clubs and Organizations associated with the Research and Development of Clinical Treatments

Logo of the Journal for the American Chemical Society

American Chemical Society (ACS)

The Georgia Tech Chapter of the Student Affiliates of the American Chemical Society is a group that has a variety of programs for interested chemistry undergraduates. The activities that are offered by the ACS range from developing professional skills and collaborating with faculty to doing cool chemistry experiments. The ACS allows for all undergraduate students who have an interest in chemistry to branch out and find what they are interested in. The ACS has research panels and networking events that help people kickstart their research into the vast world of chemistry. Aside from developing professional skills the ACS also offers fun activities every year to encourage people to be interested in chemistry and its applications. Some of the fun activities that they have held in the past include making ice cream with nitrogen and making fidget spinners using gallium. The group also attends science festivals in and around Atlanta to spread their chemistry knowledge.

American Institute of Chemical Engineers (AIChE)

The Georgia Tech Chapter of the American Institute of Chemical Engineers is just a small chapter of the biggest professional organization for chemical engineers in the world. AIChE is a large organization that allows students to develop their skills, network with professionals, and stay up to date on relevant developments in the field of chemistry. The Georgia Tech Chapter specifically organizes events throughout the year to foster a community with their members. Some of the events they plan include Lunch and Learns, social events with professionals, and guest lectures. Most of their events are more for developing professional skills and finding job or research opportunities for undergraduate students. The AIChE also hosts a lot of fun competitions throughout the year to motivate students into thinking like a chemical engineer. For example, they host a ChemE Jeopardy contest and a ChemE Car challenge where students have to build a car that runs completely off of a clean chemical source.

American Medical Student Association

Going into the medical field can be a daunting and seemingly impossible mission, but organizations such as AMSA make going into the medical so much more accessible and almost user-friendly. AMSA offers not only several pre-health workshops surrounding applicable medical skills - such as suturing, CPR, and other medical skills - but AMSA also offers a social aspect to prospective medical students. The organization has chapters in many colleges and universities, and the Georgia Tech Chapter is no different in the amount of resources and outreach offered. The organization holds fundraisers for several non-profit organizations around the Atlanta area, as well as hosting mentor-student relationships between pre-med underclassmen and upperclassmen, who have experience applying to medical school and going through the prerequisite classes important to the medical field. The organizations also holds other events, such as study sessions or yoga or even trivia night. The largest event hosted by AMSA at GT is the pre-health conference, which features pre-med workshops, several medical school representatives, as well as a keynote speaker and a panel. AMSA offers a connecting into the medical field that often is elusive to undergraduate students, and the organization helps facilitate turning pre-med students into actual med students.

Biomedical Engineering Society

Research and development might initially appear to be difficult, especially by yourself. Luckily, there exists many communities of dedicated researchers to join. One such community is the Biomedical Engineering Society (BMES). Their mission is to “promote a collaborative and inclusive community to advance human health through education, discovery, and translation”. Established in 1968, is the professional society for biomedical engineering. They currently have more than 6,800 members sharing a common vision “of health and wellness for all through engineering innovation and a steadfast commitment to diversity, equity, and inclusion”. Members also have access to BMES’ 2,000 scientific presentations, exhibit hall, and sessions of networking and career development. Additionally, members receive a monthly newsletter and subscriptions to four journals. The BMES can offer all these benefits through partnerships with the Federation of American Societies for Experimental Biology and the American Institute for Medical and Biological Engineering. By joining professional societies like BMES, one can increase the resources you can access as well as the opportunities to develop lab experience and your resume.

Healing through Autoimmune Disease

A wellness center that aids patients for holistic treatments through integrative whole-body therapies and by focusing on nutrition consultations. It is a center for individuals with autoimmune disease since treatments are rare to work with and provides alternative health outside of lab medicine. The REGN 7257 chemical being used as treatment is very new in the stage of clinical trials and its effectiveness is unknown to patients struggling with aplastic anemia, so this center is just an alternative route not even for these specific people but those with low treatment options. That usually occurs for people that have autoimmune systems since their defensive line in their own body cannot do enough to sustain them their entire quality of life and treatments are rare to fix that. So, for motivation and an alternative option this center acts as an extra chance or a chance in between treatments which REGN 7257 is currently in. It is in between clinical trials and what could be a genuine treatment option.

IPSO (Intellectual Property Student Organization)

The intellectual property student organization is a charter at Georgia Tech that is geared towards allowing students who are interested in pursuing a career in intellectual property, or a career that is adjacent that would benefit from this knowledge. This organization helps those who are pursuing a variety of careers from engineering to digital design to understand the network of intellectual property and how it would be a crucial aspect of their future careers. One of the main features is that this community is not limited to those studying public policy specifically, and is open to all majors who want to participate. This organization also serves as a basis to meet professionals in fields such as consulting, law, and business and help students learn the applications and importance of understanding this legal doctrine. Additionally, this is a newly rechartered organization and is geared towards creating a community that is made from both Georgia Tech students and professionals in the greater Atlanta area.

Microbe Club

This society offers a membership for scientists that are interested in the uses of microbes and the way they can be applied to practical uses. It is a large microbiology society based in Europe that also extends its memberships across universities and research institutes. In most recent findings they studied the cases of the drug resistant bacteria Shigella and spread light about research in that realm. Since microbiology is the study of organisms too small to be seen by the naked eye, autoimmune diseases pop up in conversation since individuals struggling with this disease are so prone to infection by bacteria or viruses. Basically, the society gives a forum to microbiologists and scientist that want to indulge in a microbiology career while holding conferences as well to discuss the influence of their works. The journals held by the society hole work from general virology to medical microbiology which is a realm that the diagnosis and symptoms of aplastic anemia would end up in. Their articles are open access to the reader after publishing but the ones that wish to publish are only those that are a member.

Patent Pro Bono program for inventors and small businesses

This organization is sponsored by the USPTO to help smaller inventors and companies mitigate the financial burden pursuing a patent could bring. Since this process costs hundreds of thousands of dollars to acquire and manage, this social program is geared towards allowing equal opportunity for all inventors to have that same right to own their invention. In order to be a part of this program, the household income of the inventor must be less than three times the federal income poverty level. Additionally, this program only takes into account people who have already submitted a probationary patent or have a pending patent with the USPTO. The reason is, on top of the patent application itself, the legal fees required to negotiate the patent claims are also costly. This program is a base for both inventors who need the pro bono litigation help, and lawyers who choose to volunteer their time at this organization to help those acquire ownership of their invention.

Student Hospital Connections at Georgia Tech

For a lot of undergraduate students, gaining experience in the medical field in preparation for medical school is often a challenge. The Student Hospital Connections at GT (SHC) is an organization that facilitates clinical opportunities for pre-med students at Georgia Tech. Several different medical institutions in Atlanta are connected with the organization, allowing students to gain valuable mentorships and even references for their future in the medical field. SHC also has a reimbursement program to assist students with costs associated with traveling for medical experience. The organization has formed valuable with many medical centers throughout Atlanta, which allows involved students to gain valuable medical and clinical experience in preparation for entering into the medical field. The organization also acts as a service organizations, volunteering with several other care-oriented organizations throughout the Atlanta area, benefiting members with valuable experience, and helping the community through service. SHC also holds other events and panels to assist with pre-med advising and how to navigate the pathway to enter the field of healthcare.

Undergraduate Research Ambassador

While the research and development involved in creating new drugs like REGN 7257 might seem lightyears away, Georgia Tech makes sure to grant you the opportunities to gain on-hand experience in a laboratory setting. One way this is achieved is through their Undergraduate Research Ambassadors (URA). Through this program, peer undergraduate researchers help mentor other undergraduates navigate the research process. This includes seminars explaining the ins-and-outs of emailing a lab PI or scheduling and conducting meetings with graduate researchers. URA also maintains a database of all available research opportunities on campus to help connect students to new passions. Additionally, URA also hosts events like the Undergraduate Research Symposium and casual mixers to give undergraduates as well as researchers the opportunity to meet each other without the stress of an academic setting. The program is filled with passionate undergraduates eager to share their experiences and joy of research to others. In this way, URA maintains a dedicated community empowered to develop their own undergraduate research program to facilitate this mentorship.

Doctors Without Borders

DWB logo in french

The main problem with diagnosis of Aplastic Anemia is that the services that are needed to diagnose Aplastic Anemia often aren't available to the less fortunate. Doctors without Borders is able to alleviate this deficit. The objective of the Georgia Tech chapter of Doctors Without Borders is to aid Doctors Without Borders (DWB) by adopting a global outlook and taking action within the local community. This involves educating the campus and Atlanta communities about the healthcare crises faced globally, the work of DWB, and how individuals can contribute to the cause. It also entails raising funds to support the work of DWB and volunteering with organizations that support DWB's mission and/or work towards alleviating health disparities. By fostering awareness and promoting action at a local level, the organization seeks to make a meaningful impact on global health issues and support the efforts of DWB.

American Association of Medical Colleges

One of the clubs on campus that allows students to potentially witness the procedures being done to confirm the diagnosis of Aplastic Anemia is the American Association of Medical Colleges. The AAMC is a non-profit association that was established in 1876 and operates out of Washington, D.C. Its primary mission is to lead and serve the academic medicine community in enhancing the health of individuals globally. The organization is dedicated to achieving this goal through various initiatives, including medical education, healthcare, medical research, and collaborations with the community. By focusing on these areas, the AAMC aims to bring about transformational change in the field of healthcare and to contribute to the betterment of health outcomes worldwide. Through its ongoing efforts, the AAMC seeks to create a brighter future for individuals across the globe and to build a world where everyone has access to quality healthcare services.

Glossary

Bone Marrow Transplant

Bone marrow transplant is a procedure used to replace defective or faulty bone marrow in patients that cannot produce enough healthy red blood cells. Transplants can either be allogenic - from a donor, or autologous - from a separate area in the body containing healthy marrow. Bone marrow transplants are used to treat many different conditions, including different cancers such as leukemia and lymphoma, syndromes and disorders including aplastic anemia and bone marrow failure, and many other applicable diseases. In a bone marrow transplant, marrow from the donor (either a healthy marrow site in the body or donor tissue) is collected by filtering stem cells from the blood. If marrow comes from a separate donor, a sample of bone marrow may be used instead of stem cells from the blood, dependent on which is a best fit for the transplant recipient. These stem cells are then frozen and prepared for transplants, while the remainder of the blood is filtered back into the body. To receive a transplant, conditioning of the current marrow and body must take place, in order to negate infections or complications caused by bacteria or viruses that could be present. This conditioning often includes different types of chemotherapy or radiation, or in some cases immunosuppression. During the transplant, donor stem cells/marrow are either injected into the blood stream through a central line, or delivered directly to the area containing faulty marrow. The new stem cells then integrate into the defective marrow and begin producing red blood cells. As the stem cells integrate into the marrow, blood infusion are common in order to maintain a healthy level of red blood cells until the morrow transplant is fully effective.

Complete Blood Count

A complete blood count is a type of blood test that can be used in diagnosis several different conditions, including anemia, leukemia, or generalized infections. In this test, a sample of blood is drawn, usually via an IV located on the inner elbow. The blood sample is then sent to the lab for analysis. The complete blood count measures the amounts of different cells located in the blood, including: red blood cells, white blood cells, hemoglobin (oxygen-carrying cells), hematocrit (percentage of blood made up of red blood cells), and platelets. In general, a CBC does not necessarily lead to a concrete diagnosis of a condition, but the test does provide vital information that can narrow down possible diagnoses. Different results from the test can indicate different conditions. For example, if white blood cell count is measured abnormally low, this would indicate some sort of issue with the immune system such as an autoimmune disorder. If the white blood cell count is high, this indicates that some sort of infection may be present. Using the information from a CBC, physicians are able to narrow down a broad scope of diagnoses, and see more specific or hidden symptoms that may not present with the patient's physical symptoms.

Cytokine Receptors

Cytokines are secreted regulatory proteins involved in extracellular signaling throughout the body. They are produced by a variety of cells with cytokine production being found in many producing cells. Cytokines can effect many cell types provided they express the correct receptors. The production of cytokines is transient but pleiotropic; they act over a short distance but affect a many different cells and tissues. While the many cytokines have differing structures, different cytokines have overlapping actions. It should be noted that these properties are a generalization of cytokine receptors with many outliers existing. REGN 7257 takes advantage of the common cytokine receptor γ-chain (γc) of the interleukin-2. Produced by a type of T lymphocyte, the cytokine increases the growth and activity of surrounding T and B lymphocytes. The new drug’s immunosuppressants will attempt to block use of the cytokine by having cytokine-specific neutralizing antibodies (CNAs) or soluble receptors bind to the cytokines. Blocking the IL 2 cytokine will prevent the growth and continued activation of T and B lymphocytes.

Dissociation Constant

The dissociation constant of a compound is a specific equilibrium constant that measures the affinity for a larger molecule to separate into its smaller constituents. For the sake of drug synthesis the dissociation constant, K_D, is a constant used to describe the affinity between a ligand (the drug) and a protein. This constant also tells how tightly bound the ligand will be to the protein it attaches too. This constant is affected by the certain types of bonds included within the two molecules such as hydrogen bonding, electrostatic interactions, and van der Waals forces. A small dissociation constant value means the ligand has a greater binding affinity for its targeted protein while a large dissociation constant means the ligand will weakly bind to its target. Regarding the synthesis of the REGN 7257 drug multiple dissociation constants are listed for different temperatures to show what temperature is the best for binding. All of the dissociation constants listed are extremely small for temperatures across 25^oC and 40^oC which means the antigen binds to interleukin-2 gamma the best between these temperatures.

Model of epitopes

Epitope

The epitope is the small site on an antigen in which another antibody may bind to. The epitope of an antigen is not however always in the same place and is not defined by a specific sequence of amino acids. Depending on the structural properties of the antibody (covalent bonds, ionic bond, hydrophilic, and hydrophobic interactions) there is a range of possible binding sites. The epitope of an antigen is usually the residue of one to six monosaccharides or five to eight amino acids on the surface of the antigen. Since an antigen exists in 3D space the epitope can exist either as a linear sequence or a folded sequence of amino acids. The epitope is important in the synthesis of REGN 7257 because any antibody that is used in the creation of it binds to the epitope of the interleukin-2 gamma chain. The epitope is the basis for all of the binding in the synthesis of the REGN 7257 even though the compounds are dissolved in the synthesis.

Hematopoiesis

a visual of a substance being injected into the bone marrow and what that would look like cellularly

Hematopoiesis is defined as blood cell production in order to replace old damaged ones. This formation occurs in the bone marrow and continues as an ongoing cycle in life to constantly keep a blood supply. The bone marrow makes three groups of cells that differentiate and specialize into either red blood cells, white blood cells, and/or platelets. A lack of white blood cells is what would cause an individual’s immune system to be shot and allow invaders to enter the body. REGN would focus on either increasing those counts of killer cells or strengthening the minimally existing ones for people whose bodies cannot do it on their own. But in the case of aplastic anemia, the entire bone marrow is unable to make more cells to function properly and fight diseases properly, so hematopoiesis will try and shift to the liver or spleen or even lymph nodes and continue trying to produce necessary cells from there usually by the help of a chemical substance such as the treatment described for patients with aplastic anemia.

Macrophages

They are a type of specialized cells that present antigens to T cells and activate other cells by inflaming the cells to release cytokines. They originate from blood monocytes which stem from the bone marrow but then instead of remaining in the bloodstream they differentiate in different tissues throughout the body. These cells require a high level of specialization which allows them to recognize numbers of pathogens and produce even higher levels of inflammatory cytokines to aid the immune system. Macrophages can migrate into almost every tissue making them essential for pathogen control and immune system defense. This migration also goes to tissues to the lung by a particular group of macrophages that fight respiratory pathogens. Another group also specializes to the liver called Kupffer cells that work on initiating the immune responses in the area. They detect all these bacterial products using receptors like TLRs that bind to specific pathogens and then remove them from the body.

Monoclonal Antibody

Application of Monoclonal Antibody for Fluorospot Assay

In the human immune system, foreign substances are attacked with large numbers of antibodies. These antibodies are a protein that sticks to a specific protein called an antigen. REGN 7257 is an anti-interleukin 2 receptor subunit gamma [IL2RG] monoclonal antibody. In other words, REGN 7257 is a synthesized protein made in a laboratory that can bind to certain targets throughout the body. Because the new drug is an anti-interleukin 2 receptor, it is engineered to bind to the IL-2 receptors on the surface of lymphocytes. The specific targeting nature of the monoclonal antibody overcomes the off-site toxicity plaguing conventional drug delivery methods while improving the efficiency of the drug with lower doses. Monoclonal antibodies can be made using four different methods: murine are made from mouse proteins, chimeric are a combination of mouse and human proteins, humanized are small parts of mice proteins combined to human proteins, and human are fully human proteins. While this new technology is a powerful new tool, there are some known side effects with allergic reactions and low blood pressure being the most severe.

Patent

Issued United States patent cover page

A patent is a legal document that is obtained through the United States Patent and Trademark Office. This legal document gives its owner exclusive rights that allow them to monopolize that invention for 20 years for a utility patent after the issued date and 15 for a design patent after the issued date. This right includes preventing others from creating the same product even if it is reverse engineering and created independently. Patents are not the only way to protect intellectual property, but for most scientific and technological innovations these are the most suitable just due to the nature of the industry. Patents are also self-enforced making them a pricey means to manage intellectual property. The USPTO has no responsibility to look out for inventors’ patents and prosecute if they observe an infringement. This responsibility is solely placed on the legal counsel of the patent owner and therefore patent infringements are not always pursued.

Prior Art

In order to obtain a patent, an inventor must undergo a process when the application is evaluated and thereby approved by the USPTO as something that is novel and useful to society. During this process, the patent examiner must test whether or not the product or process is novel by looking at all of the work that has been done in the field up to this point. Prior art refers to any sort of medium that relates to the work that has been published in the field. Usually, these works of prior art are used to negate the issuance of a patent since the mention or notion of the product or process in other papers could lead the examiner to deem that the invention was obvious or anticipated. Obvious applications have 2 or more independent occurrences of prior art and anticipated has one. Once patents have been published, they too become prior art and accessible to the public.

Paroxysmal Nocturnal Hemoglobinuria (PNH)

PNH is a medical condition that is a rare blood disorder. If left untreated, PNH can lead to hemolytic anemia, chronic kidney disease, and thrombosis (blood clots). Hemolytic anemia occurs when red blood cells are destroyed at a faster rate than they are produced, leading to a shortage of oxygen-carrying cells in the bloodstream. PNH and Aplastic Anemia often have very similar symptoms, so it is important to determine which disorder is causing the symptoms that the patient is presenting with.

Bone Marrow Extraction Procedure

Bone Biopsy Needle

Before the procedure, the patient's vitals are checked and they are given a form of anesthesia to help with the procedure. The area where the biopsy will be performed (usually the hip or breast bone) will be sanitized. The procedure itself involves a bone marrow biopsy needle, which is a specialized needle that is of a larger gauge. This needle contains a separate smaller needle within it that it removed before it is inserted into the patient. The larger gauge needle is inserted into the patient and rotated 360 degrees in order to separate the core sample from the rest of the bone. The large needle is then removed from the patient and the smaller needle is used to push out the sample from the larger needle to be extracted.