Custom antibody production is a laboratory activity that has attracted the attention of many scientists. With technological advancements, large-scale production of antibodies is now possible. Antibodies play an essential role in fighting viruses. Custom antibodies are vital for research and laboratory experiments. They are produced by immunizing rabbits and mice with specific antigens.
The immunized animal synthesized the required antibody, later retrieved through cells. The antibodies must undergo purification. The purification process helps eliminate unwanted components such as antigens before immunization. This article will provide four secrets to antibody production.
What are antibodies?
An antibody is responsible for immunity and fighting diseases in living organisms. They respond to any invasion of the body by foreign molecules. Structurally, antibodies are Y-shaped and are made up of polypeptides, heavy chains, and light chains. A custom antibody is manufactured in the laboratory by immunizing a specific animal with an antigen. The antigen induces the production of antibodies in the animal, later retrieved.
If you consider producing a custom antibody, the following secrets will be of good help.
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Animal selection
The custom antibody is produced from a laboratory animal immunized with a specific antigen. Therefore, the selection of an animal is significant in antibody production. However, before selecting an animal, you should first choose the antigen you require and prepare it adequately. Custom antibodies are produced from specific antigens depending on the type and usage of the antibody. For polyclonal antibodies, rabbits are the preferred animal. This is because rabbits are easy to handle.
Their size can produce more antibodies and have a high titer. In addition, other factors such as housing environment, availability of different antigens, and animal diets also affect the production of polyclonal antibodies. Before using a rabbit, you can house it in a laboratory where you can control it and place strict barriers to the environment. You will obtain higher yields of polyclonal antibodies. Housing your rabbit before using immunization will help reduce the animal’s stress level and induce a more natural behavior pattern.
For the monoclonal antibody, mice are popular and the most preferred animal in the production process due to their ability to produce hybridoma cells. Other animals such as birds, humans, and rabbits can be used. Two methods can produce monoclonal antibodies: immune antibody library and classical hybridoma fusion. The method you choose to have the monoclonal antibody will affect the type of animal you select.
Immunization process
To induce the production of antibodies in an animal, you have to immunize it with specific antigens. However, how the process of immunizing the animal will be affected by several factors such as the particular characterization of the adjuvant, the type of the animal, and the composition and volume of antibody you intend to use. In rabbits, multiple subcutaneous injections on the back are the most common immunization procedure. This protocol of immunization allows the potential inflammatory reactions to be visually monitored.
Once you have immunized the animal, you should monitor it daily. The monitoring involves examining the injected part to check on the general development of potential hazardous lesions and observing the animal’s general health. In addition, you should also monitor the actual antibody response. Fourteen days after the first immunization, you should perform a bleeds test. You can also measure the levels of specific antibodies.
Collection of antibody
Once you have observed an appropriate antibody response after immunization, you can now collect the produced antibody in the serum or the hybridoma cells. Different species of animals allow a unique blood circulation volume percentage. The production bleeds, which are the higher volume bleeds, are circulated out over time. The collected antiserum is first pooled and centrifuged to eliminate the cellular debris before testing the quality assurance.
The produced antibodies are collected directly in the serum for the polyclonal custom antibodies. In contrast, in monoclonal antibodies, the immunized animal’s secreting cells combine with the myeloma cells to create monoclonal hybridoma cells lines that synthesize the specific antibody in the cell culture supernatant.
Purification of the antibody
After producing custom antibodies, the purification process is crucial. The purification process helps eliminate unwanted components such as cellular debris, antigens, non-specific immunoglobulin, and any microorganism affecting the antibody’s quality. You should choose the appropriate purification method. Chromatographic matrix is the most preferred method for purifying polyclonal custom antibodies.
Using this method, you can conduct antigen affinity purification alone or with protein. There is also a recent development of resins that helps purify several types of antibody species, enhancing the removal of contaminants such as proteases. You should choose an appropriate antibody purification method by considering the process and the animal you have used to produce the antibody. The purification method you choose will affect the quality of the antibody.
Wrapping up
You can consider using an adjuvant to boost the results to produce high-quality antibodies, especially when using antigens with low immunogenicity. An adjuvant helps synthesize an immune-stimulatory signal that stimulates the immune response in the immune system. Once you have produced and purified your custom antibody, you can classify and characterize them. Custom antibodies can be helpful in the research and production of vaccines. Several laboratories offer quality custom production services, and you can always reach out when in need.
