Antibody Generation: 6 Things You Need To Know 

Crystal Hemoglobin Structure for anibody generation

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When a foreign particle such as a germ enters your body, the immune system responds by producing a host protein called an antibody that helps protect the body from foreign invasion. Foreign disease-causing organisms are called antigens, and the body produces antigen-specific antibodies that only bind to specific antigens and prevent infection. 

The antibody-antigen relationship is much more complex, but it is critical in fighting infection. Scientists can copy the immune response to antigens and produce antibodies of interest to diagnostic applications and research. Antibody production is a novel application that has revolutionized the understanding and management of diseases. But producing antibodies that are of biological importance is a complicated process. But with the advancement in technology this has become relatively easier. 

Many stages are involved in producing antibodies, depending on the required outcome. A mistake in any of the stages could lead to antibodies not specific to the target antigen. Do you want to know more about antibody generation? 

6 Things You Need to Know About Antibody Generation 

You Must Select the Right Animal Species. 
 

Generating polyclonal antibodies relies on the use of live animals, and scientists need to select the animals used in the process carefully. While many animals have similar immune systems to humans, not all can produce useful antibodies for research and therapeutic application. 

When selecting the right type of animal for antibody production, you need to consider the following: 

  • Number of polyclonal antibodies needed 
  •  How easy it is to obtain blood samples 
  •  The polygenetic connection between the selected animal species and the antigen 
  •  How do you intend to use the polyclonal antibody 
People Grooming a Guinea Pig

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In most cases, scientists require a few antibodies for research and clinical application. In such cases, the common animals for these studies are: 

  • guinea pig 
  •  mouse 
  •  rat 
  • rabbit 
  • hamster 
  • goat 
  •  sheep 
  • chicken 

Rabbits are often the choice animal for polyclonal antibody production because they are relatively small, are easy to handle and bleed, have a long life span, have precipitating antiserum, high affinity, and high titer production. 

If there is a need for large amounts of antibodies, larger form animals such as sheep, goats, and horses can be used. Due to animal welfare concerns, you need to consider the ease of bleeding when selecting the animal species. When there is no need to use a specific animal species, you should use the animal from which it is easy to obtain blood samples. 
 

It’s Important to Consider Antigen Purity 

One of the significant concerns with antibody production involves the purity of the antigen used. You have to consider antigen features when producing antibodies. The quality, quantity, and production process of the antigens will determine the outcome of antibodies, and all these features must be considered before the production process begins. 

Immune system specificity determines the quality of the antibodies you obtain during antibody production. This outcome depends much on the purity of the antigen. If the antigen is not pure, you may end up with antibodies that are more reactive to the impurities than the target antigen. As a researcher, you must undertake extensive antigen purification to avoid unwanted antibody reactions. 
 

You’ve to Select the Right Adjuvant. 

Sometimes, the immune system fails to react to the antigen once evoked. If this happens, it won’t be possible to obtain antibodies since the immune system will not produce any. In such a case, you may have to stimulate the immune system to respond as desired. 

Set of various medical equipment for successful taking of vein blood

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The induction of the immune system requires the use of adjuvants to direct the immune system to react to the antigen. There are more than 100 adjuvants used in antibody production, but these are the most common: 
 

  • Freund’s complete adjuvant (FCA) 
  • Freund’s incomplete adjuvant (FIA) 
  • Montanide 
  • Aluminum salts 
  • TiterMax™ 
  • Quil A 
  • RIB 
  • Iscoms 

Among these options, FCA is the most popular because you can induce high antibody titers to various types of antigens with it. 

Injection Protocol Matters 

Once you have settled on the animal species and the adjuvant, you must understand the injection process. The main thing you need to know about the injection of the antigen is that it’s highly dependent on the type of animal and the adjuvant you’ve chosen. 

Syringes are frequently used in clinical medicine to administer injections, infuse intravenous therapy into the bloodstream, apply compounds such as glue or lubricant, and draw/measure liquids.

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There are many factors to consider in the injection protocol, such as: 

  • Route of injection 
  • Volume of injection 
  • Number of injection sites 
  • Booster injections 
     

You should Monitor the Animal for Side Effects. 

Once you’ve injected the animal with the antigen, you should monitor it daily for post-injection side effects. You should also examine the animal at least three times per week to make significant conclusions about the response. 

Special consideration should be given to the injection site as there could be pathological outcomes not captured in clinical results. You should palpate the injection site to help assess the side effects. 
 

You should Monitor Animal’s Antibody Response 

Antibodies take time to develop, and the response could even take weeks depending on the antigen and type of animal. But this does not mean that you just leave the animal and wait. Instead, you should monitor how the animal responds to the antigen. 

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You should collect blood serum regularly to monitor the presence of antibodies. This process is invasive unless you’re using a chicken to collect the samples from the eggs. When collecting blood, ensure that you safely collect a safe volume. 
 

Conclusion 

Polyclonal antibody collection is a rigorous process, and you must follow strict guidelines to get high-quality antibodies. If the antigen is contaminated, you may result with antibodies that target the impurities rather than the antigen and thereby be useless. 

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