How to Make Polyclonal Antibodies in Laboratory -An Overview

Antibodies are part of the body’s defense mechanism to combat foreign material (antigens) when they enter the body.

The B cells are responsible for the production of antibodies. Antibodies are of great value in medicine, and they are commercially in demand.

Production of antibodies is a growing field among the most common pharmaceutical ventures.

The polyclonal antibodies are in higher demand since they work best against polyvalent antigens. Apart from that, they form the easiest implementation against various epitopes of antigens.

But researchers are also interested in polyclonal antibodies for their medical research and diagnosis of infections.

The production of this type of antibodies becomes a necessity for medical researchers. And you will want to know how to make the antibodies in your lab to complete your projects.

This is the process to follow to collect the antibodies you need.

1. Get Your Specimen and Immunize It

The first stage is the introduction of an antigen into a species you want to use for the antibody production project. The animal you are going to use becomes the host for the antigen.

This is called “Prime immunization”, and you will need to introduce adjuvants alongside the antigen. The adjuvants will help the host animal to respond to the antigens so that it starts the antibody production against the foreign material.

Although the most common method of immunization is through injection, some involve oral and intranasal administration.

With the recent developments, DNA vaccination is also viable, and it is the best option for complicated antigens. The first vaccine activates the B lymphocytes, which divide and become the plasma cells.

They then produce antibodies in large numbers in the bloodstream and the lymphatic system.

Depending on the amount and maturity of antibodies you need, you may need to give the animal a boost vaccine.

This helps the immune system to retain the memory of the antigen and continue producing antibodies. Otherwise, the immune system will “assume” the problem no longer exists, and the production of antibodies stops.

2. Measure the Immune Response

The body may take some time before it populates with the antibodies against the antigens you introduced into the animal.

To evaluate the progress of the immune response, you should measure the presence of antibodies regularly. Using an ELISA test, you can tell how much the immunizations have helped the specimen to generate the antibodies you would want to get.

You need serial sample dilutions to determine whether the antibodies are ready for harvesting to proceed with your projects.

Typically, you would need a 1:64,000 titer. The ratio means that the immune response against the antigen was successful.

However, a lower ratio means that you need to proceed with the boost vaccine. You can continue to boost until you reach the titer you need.

3. Harvesting the Antibodies

To complete your custom antibody production, you should harvest the polyclonal antibodies from the host animal’s system.

The harvesting is quite the easiest part of the process. You begin by drawing blood from the animal once the antibody titer reaches your desired level.

The blood contains everything from blood cells, sugars, to various types of proteins in the body. You will need to remove those substances and remain with serum.

It is the serum that contains antibodies. Centrifugation and cooling are the main stages of obtaining serum from the whole blood.

4. Purify the Antibodies

Purification of antibodies is the extraction of the polyclonal antibodies from the serum. Like other forms of proteins, there are various antibody purification methods you can apply.

Your goal is to ensure that you get the highest yield of antibodies from the serum, and, possibly, using minimum resources and the shortest time.

5. Fragmentation of Antibodies

It is advantageous to produce polyclonal antibodies because they are easier to make and harvest. They also include variations that offer different ways of binding to antigens.

In this way, the antibodies can deal with a mutating antigen, and they can handle any variation in which the antigens present themselves.

It is, therefore, ideal to fragment the antibodies when you want to use them for various specific purposes. The fragmentation part is a final stage that involves modification to make antibodies ideal for a specific target. The preliminary is to remove parts of the antibody that do not have the binding abilities.

In Summary

For whatever research you are making antibodies, you would want to consider making them polyclonal. The entire process would take you a few weeks to complete.

And, wherever you need to boost the vaccine to enhance antigen “memory” in the host, you would need more time. You, therefore, want to have everything accurate from the beginning.

After making the polyclonal antibodies, you can label and assign them various uses including ELISA, cellular imaging, western blotting, and other antibody-use techniques.

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