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Monoclonal Antibody

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The human immune system is strange. it is such a complicated system that can’t be imagined. It is a well-developed process in the human body. Whenever any foreign element inserts into the human body, the immune cells start acting. It starts operations on the foreign elements & destroys them within the body. It is a complicated structure. Whenever there is any threat from the foreign element, it is considered as the antigen. The element that is not a part of the body. And the substances that help to destroy the antigen are known as the antibody. The monoclonal antibody is one type of that.

Definition of Monoclonal Antibody

A monoclonal antibody is a type of antibody that can able to operate against only one type of antigen. That is why it is called the Monoclonal Antibody. The word “Mono” means that the antibody can able to interact with only one antigen. The short form of this antigen is the Mabs or Moabs. This antibody is only able to create artificially. This means this antibody is made up in the laboratories.

The human body often comes across different foreign elements. While interacting with these foreign elements, there might be some fever in the human body. To destroy those foreign elements, human bodies start producing monoclonal antibodies. Now, those monoclonal antibodies are taken from the human body. Then in the laboratory, the number of antibodies is increased with some processes. The medicine for such disease is now ready in the market.

Monoclonal Antibody


Structure of Monoclonal Antibody

The structure of the monoclonal antibody is one of the simplest structures. The structure of the monoclonal antibody is a “Y” shaped structure. This is entirely made up of proteins. This structure can’t be divided further structure. But there are two chains present. these chains are the protein chains. And these have structural differences.

  1. Heavy Chain: This is the lower part of the monoclonal antibi. This is the part where only a vertical chain is present. this chain is known as the Heavy Chain. As this chain can’t able to move or do any processes. This chain is the indicator that the antibody has successfully attached to the antigen. After successfully attaching to the antigen, the heavy chain is only exposed. So, there is no change to remove the antibody from there. This is known as the constant region. As there is no need to change this part with different foreign elements.
  2. Light Chain: Light chains are the main player of the monoclonal antibody. This is the upper part of the monoclonal antibody. There is a total of four arms in these regions. On both sides, there are two arms. These arms help to bond with the antigen. Only with a single arm, the antibody can’t able to bond with it. This region is known as the variable region. As these region needs to be changed with different antigens. Only a special type of antigen can bind with this spot. Different antibodies are required for different antigens. There are hinges in the connecting area of those two pairs of bonds.

Production of Monoclonal Antibody

It is not always possible to create monoclonal antibodies in the laboratory from a sample taken from the human body. Many serious diseases in the world can not be intentionally infected by any human being to get monoclonal antibodies. In these cases, scientists take the help of mice. Suppose, there is a need to create a monoclonal antibody for any foreign element. For that purpose, there is a need to have the monoclonal antibody that is created inside of anybody. Then using that sample more antibodies can be produced in the laboratory.

So for that reason, the foreign element is inserted into the body of any mouse. So, the mouse is now affected by that disease. Now, the mouse body starts making the monoclonal antibody. Now, the cells that are creating the antibody need to be taken out of the mouse body. Mainly these are the spleen cells. now, in every higher animal body, there is a special cell found. These are known as Myeloma Cells. These cells are also taken out of the body. Now, in the laboratory, they are diffused. After fusion, new artificial cells are created. These are known as Hybridoma.

Now, there must be lots of cells in the Hybridoma. All of these cells might or might not create the certain antibody which is looking for. Also, sometimes among those cells, any one cell is creating purer & best antibodies among all of them. so, the task is to identify the best cell from the group of Hybridoma. For that reason, now the cells are exposed to culture in the HAT medium. This is a special kind of solution or medium that is being used to identify the best Hybridoma or the positive cell. Now, after identifying the best cell, it is harvested. Now, the monoclonal antibody is created in a large volume. So, it is now ready to start operation on any human body.

Hybridoma Technique

In the process to develop the monoclonal antibody. One of the important steps is to develop Hybridoma. And as per the name of the developed cell, the process is known as the Hybridoma. At first, the antigen is inserted in the body of the mouse. Then it left for 72 hours. After 72 hours of the injection, the spleen cell of the mouse is collected. Spleen is the most important cell in the immune system. in the spleen cell, the antibody from the antigen develops. So, the spleen cells are taken for developing the monoclonal antibody.

Now, one of the major disadvantages of these cells is, they are not immortal. This means after taking out those cells for some time those cells will have died. But the intention is to create the immortal version of those cells. for that purpose, there is a need to fuse with myeloma cells. Myeloma cells are cancerous cells. This means they will always develop until there is a disconnection of the nutrients. Now, the intention is to fuse the B cells of the spleen & the myeloma cells of the mouse.

These two cells are placed in the same medium for 48 hours. They are placed in a medium named Polyethylene Glycol.  In this medium both the cells are placed to fuse. After the fusion, a new cell will be derived. The cells will have the antibody production power from the B cells of the spleen as well as they will be immortal as the myeloma cells. These cells are termed Hybridoma. This means these cells are the hybrid form of the cells. and this overall process is known as the Hybridoma process.

HAT Medium

Hybridoma Technology


It is one of the essential elements in the production of Monoclonal Antibodies. This medium is completely used to extract the hybridoma cells from the mixture of the medium. The HAT medium is made up of three components. They are the Hypoxanthine, Aminopterin, and Thymidine. The first letter of these elements is being used to name this medium. The H from the Hypoxanthine, the A from the Aminopterin & the T from the Thymidine. All these substances are combinedly and named the HAT medium. All these elements are proteins. And their main goal is to stop the growth of any cells in the proper medium. This medium is used in the production of the monoclonal antibody.

The HAT medium is not alone able to do the entire task. This means the cells can’t able to be placed in a single HAT medium. The cells where it is from the beginning of the experiment, there the HAT medium should be included. Hypoxanthine is the substance that deactivates purine protein in a certain medium. Purine is a type of protein that is used in the growth of cells. This is the protein that is being used as a nutrient for the growth of any cells. Aminopterin is a type of drug. That helps to stop the metabolism of any cells. This means the cells can’t able to grow more in the medium. And Thymidine creates issues with the synthesis of the DNA in any cell. This means all these elements participate to stop the growth of the cells at any cost. The influence of these substances can only recover by the Hybridoma cells.

As the HAT medium can kill natural cells, the cells that are not introduced by the nature can skip this effect of the medium. Purine is the protein that helps to make the growth of cells. now, this protein is created using some special pathway. They are the De Novo pathway. For creating some proteins, there are certain processes. They are the list of processes. Now, the Purines can able to synthesize using the De Novo pathway. And in absence of the De Novo pathway, the Purine can be produced using the Salvage pathway. Now, the components of the HAT medium create an issue in the formation of the Purine proteins.

The Aminopterin substance in the HAT medium at first blocks the De Novo pathway of all the cells present in the medium. So, the pathway can’t able to use by the cells to produce the proteins. So, by force, they have to follow the Salvage pathway. In the Salvage pathway, two essential elements are needed. One is a special type of enzyme & another is a protein. The enzyme is the Hypoxanthine Guanine Phosphoribosyl Transferase (HGPRT). And the protein is the Thymidine Kinase. The Thymidine Kinase also acts as the enzyme there.

Now, the Thymidine Kinase is blocked by the Thymidine in the HAT medium. So, the cells can’t able to access that element for life long. And unfortunately, there are no HGPRT enzymes present in the Myeloma cells. So, as a result, the Myeloma cells can’t able to live more in this medium. As there is no HGPRT key element present. So, as a result, eventually, all the Myeloma cells will be destroyed. Now, there are only the Hybridoma cells & the spleen cells present.

The Spleen cells can able to replicate here. As they have the HGPRT present in their cells. But as they are not Myeloma cells. This means they are not ever-living cells, so eventually, they have to die. So, with time, the spleen cells will be removed from the medium. Now, the Hybridoma cells will only survive there. As Hybridoma cells come from nature, they have some special features. As they have derived from the spleen cell, so they can create the HGPRT enzyme in their cells. And as they are a hybrid with Myeloma cells, they have the feature of cancerous cells. This means they will never live.

So, the HAT medium is used to remove all the unnecessary cells. The Myeloma cells need to have died as there is no option to create an HGPRT enzyme. And spleen cells need to have died as they are not ever-living cells. But they have the HGPRT enzyme. As the Hybridoma cells have all the features so they will only survive in this medium.

Purification Technique

Purification is the second most important process for developing the monoclonal antibody. After the hybridoma process, there is a mix of cells in the hybridoma solutions. As all the spleen cells & the myeloma cell will not be fused. Among the quantity of them will be fused & some of them will not be fused. So, in the solution, there will be three types of cells present. One is the newly created Hybridoma cell, another will be the B cells of the spleen and the last one will be the myeloma cells.

Now, the main goal is to only extract the hybridoma cells. for that purpose, all other cells need to be eradicated. For that purpose, all the mixture is placed into a new solution. That is the HAT solution. This is a special solution. In this solution, there are three substances present. They are the Hypoxanthine, Aminopterin, and Thymidine. These substances will act on the myeloma cells. These substances will cut down the nutrients supply for the myeloma cells. in simple terms, the myeloma cells can’t able to grow more or live more in this solution. So with time, all the cells will be destroyed.

Now, two cells are remaining. One is the hybridoma cell & another is the B cell of the spleen. There is no need to do any experiments on the B cells. As they are not immortal. With time, these cells will be destroyed automatically. So, there will be simple the Hybridoma cells will be present. And that is the goal of this process. This overall process is known as the Purification process. As the other cells will be removed from the mixture.

Importance of Monoclonal Antibody

Monoclonal Antibody is a milestone in medical science. A monoclonal antibody helps to develop medicines for diseases. From the normal cold cough to some critical diseases, the monoclonal antibody is being used. The monoclonal antibody consumed very less amount of cost. So, they are pocket friendly. For a wide range of use, the monoclonal antibody is one of the essential elements in medical science. These antibodies can able to increase the immune system of the body before infecting with the disease. The monoclonal antibody is being used to treat some difficult diseases. So, there is a huge importance of monoclonal antibodies that is can’t be expressed in terms.

Functions of Monoclonal Antibody

  1. Diagnostic Testing: Monoclonal Antibody is being used for the diagnostic purpose. As these antibodies are made to act on special substances, they also help to find it out. In the testing, the antibody is used for a particular element in the blood vessels of that patient. If the antibody is attached to that substance, the result can be seen in the test report.
  2. Medical Uses: Monoclonal Antibody is used in various medicines. From the natural cold cough to some viral infection, the monoclonal antibody is being used all the time. Doctors prescribe antibacterial capsules for the treatment. There is a monoclonal antibody present for better treatment. So, the medical purpose is the main objective of the monoclonal antibody.
  3. Pregnancy Testing: Monoclonal antibody is often used to do pregnancy testing. The monoclonal antibody can find out the Human Chorionic Gonadotropin hormone. These substances are available in pregnancy testing kits. So, this antibody indirectly helps to find out the pregnancy of any woman.
  4. Treatment Of Cancer: Scientists and doctors try their best to find out the monoclonal antibody of cancer. The main need is to develop the antibody that will attach to the antigens of cancer. Hence, the cancer cells can’t able to develop more. And this will be an easier path than chemotherapy.
  5. Pain Remover: The painkiller injections that are being used by doctors in the severe case of pain, there is the presence of the monoclonal antibody. This injection helps to reduce the pain for a certain cause immediately. Monoclonal antibodies are being used in those injections. 

Side Effects of Monoclonal Antibody

As the monoclonal antibody is the protein element. And inserting more proteins from the outside of the body can cause allergic reactions. As there is a protein balance in the body. If there is a problem in the protein balance, then there might be some issues. There are lots of side effects of a monoclonal antibody. A person may feel the following issues as a side effect.

  1. Fever
  2. Weakness
  3. Headache
  4. Rashes in different parts of the body
  5. Vomiting
  6. Nausea
  7. Diarrhea

FAQs on Monoclonal Antibody

Question 1: What is a monoclonal antibody?


A monoclonal antibody is a special type of antibody, that can able to work against only one antigen. This antibody is created in any animal body for the first time. Then the sample monoclonal antibody is doubled in the laboratory using different processes.

Question 2: How many protein chains are there in the monoclonal antibody?


In the monoclonal antibody, there are two chains are present. On the lower side of the antibody, there is the heavy chain. And in the upper side of the antibody is the light chain.

Question 3: Which chain acts as the antigen receptor site in the monoclonal antibody?


The Light chain is being used as the antigen receptor side. The arms in this region help to bond with the antigen receptors. This is the most important area.

Question 4: Which chain doesn’t perform any significant operation in the monoclonal antibody?


The Heavy chain doesn’t have any significant operation in the monoclonal antibody. Their function is to provide one stable figure to the antibody. Also, this is being used as the indicator in the monoclonal antibody.

Question 5: How many arms are there in the light chain of the monoclonal antibody?


There is a total of four arms are present in the light chain. On each side, there is one pair of arms present. Among the arms, one is a big length & another is a relatively small length. These arms help to bind with the antigens. One single arm is not efficient for bonding with the antigens.

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Last Updated : 02 Jan, 2023
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