How Polyclonal Antibodies Are Produced

How Polyclonal Antibodies Are Produced 

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Early detection is very crucial in the management of many diseases and infections. Advanced technologies have skyrocketed in the medical world to pick out a specific indicator of an illness. One of these MedTech inventions is the production of polyclonal antibodies.  
 

Unlike monoclonal antibodies, polyclonal antibodies (PAbs) are a population of immunoglobulin molecules from multiple B cell lineages. Therefore, each antibody in this group reacts to a different site of the same virus, bacteria, or another molecule, collectively called antigens or pathogens.  
 

A better improvement in polyclonal antibody production is the custom polyclonal antibody production that involves generating antibodies from the adaptive immune system in response to specific offending pathogens. These antibodies have a lock and key recognition pattern for the pathogens in question, making it easier to detect an infection. 

 

Polyclonal antibody production is cheaper, quicker, and more convenient and involves a few steps. 

Production Of Polyclonal Antibodies 

 

To generate polyclonal antibodies, an animal receives an antigenic injection that will trigger the immune system to release antibodies to fight the pathogen. The antibodies produced are then harvested, prepared, and stored for other purposes. There are five primary steps in the generation of polyclonal antibodies, and these include: 
 

  • Preparing the antigen  

  • Selecting and preparing an adjuvant 

  • Picking the preferred test animal 

  • Injection of sample 

  • Extracting serum  
     

Antigen Preparation 

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The antibodies produced during the procedure are antigen-specific and should be able to recognize and bind to a particular pathogen. However, the antigen selected for the process must be pure and free from toxins to achieve this specificity. Using an impure antigen will produce immunodominant antibodies that react more to the impurities than the actual pathogen. 
 

Screening the antigen extracts for any endogenous toxins is crucial to achieving antigenic purity. Also, it is essential to sterilize and clean the materials used in the extraction, handling, and storage of antigens.  
 

Generally, bigger antibodies have more epitopes or binding sites that are easy to detect. If the targeted antigen is very small, linking it to albumins or hemocyanins is vital for appropriate detection. Apart from ensuring a large antigen size, it is crucial to use techniques that do not contaminate your preparations.  

Use Of Appropriate Adjuvant. 

 

Antigens have different degrees of immunogenicity- the ability to provoke an antibody reaction to a presented pathogen. This ability should be high in antigens used in PAbs generation, and if you cannot achieve the required levels, adjuvants come in handy.  

 

A weak immunogenic antigen can elicit an appropriate antibody reaction if mixed with a suitable adjuvant. Though, the selection of a safe and compatible adjuvant is not straightforward. Some adjuvants may provoke extreme reactions that may reduce the quality of produced antibodies or harm the animal.  
 

Based on the sensitivity of the production process, the selection of adjuvants is very critical. The adjuvant picked should be able to evoke a response and have no effect or purity of antibodies.  

The Choice Of Animal 

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Picking an ideal animal depends on the number of antibodies needed, the closeness of the antigen donor and recipient antibody producer, and the characteristics of antibodies desired. The young adult rabbit is the most preferred animal due to their strong immune responses.  
 

When large amounts of antibodies are the objective, a larger animal, a horse or goat, is ideal. Also, the farther the animal species is used, the higher the antibodies’ quality. For instance, a cat and a dog may produce better polyclonal antibodies than a lion and a cat. The animals selected should be healthy and easy to handle. Unhealthy animals may be immunocompromised and produce defective or fewer antibodies. Furthermore, aggressive animals may contaminate the samples during the injection. 

Animal inoculation 

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The delivery system depends on factors like the choice of animal and the number of antibodies one wishes to collect. So, you’ll pick the route of injection, whether intravenous, intramuscular, intraperitoneal, or intradermal, depending on the safety prognosis of the animal. 
 

The other key factor to consider before the injection is the volume of the antigen-adjuvant mixture. Only threshold volume or the smallest possible quantity of antigen that can evoke the production of antibodies is necessary. Any excess may result in hypersensitivity reactions that are harmful to the animal. 

 

Also, the number of injection sites and whether to give subsequent injections are critical factors. The lesser the number of injection sites, the lesser the chances of occurrence of secondary infections. However, more points may be preferable for the injection mixture to transfer to the whole body for more antibody production. 
 

Subsequent or booster injections may determine the quality and quantity of polyclonal antibodies released. But, the animal species, size, and injection mixture composition are vital controllers of subsequent inoculation. 

Extraction Of Blood Serum 

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After a specified time after insulation of the selected animal, experts draw serum from the animal’s blood to obtain the generated antibodies. This process is a critical stage and requires intensive care. Different techniques single out and extract the antibodies of choice for other uses. 

Conclusion 

 

The production of polyclonal antibodies is relatively inexpensive and less demanding than monoclonal antibodies. Besides, polyclonal antibodies play critical roles in the diagnosis, management, and prognosis of different diseases. Based on its crucial role in medicine, its production and use are extensively growing worldwide.