Capra Science has developed its own immunization program that optimizes the production of antigen-specific IgG or IgY antibodies while simultaneously minimizing the background (the presence of low-medium affinity non-antigen specific antibodies). Some of the most important factors that we take into consideration when forming a successful immunization program to make high quality polyclonal antibodies are 1) time, 2) the immunization technique and 3) the bleed frequency.
TIME IS KEY
Regardless of species, it is not possible to rush the natural process of an immune response. If you would like to be rewarded with an antiserum containing your top-quality antibody in high titers, a few extra weeks of patience is required. The Capra Science Immunization Program™ is optimized for reaching the highest possible levels of antigen-specific antibodies with respect to the necessary immunological processes required.
Time Allows High Yields of Specific Antibodies
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The yield and the titer has a correlation. The antibody titer is the relative concentration of antigen-specific antibodies in a sample. Moreover, the titer value is the sample dilution at 50% of the max level of absorbance.
Reaching high titers depend on many parameters, one of them being time of sample collection after first exposure (immunization). Top titer-levels are often reached after 3 months (see schematic figure on right hand). |
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Time Allows High Affinity and Low Background
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A top-performing antibody or an antibody with high quality is often refered to as having high affinty to the antigen and being specific to the target.
The Capra Science Immunization programs™ are not “express” or short. The reason is that high specificity and affinity towards the target antigen can only be reached given enough time (see diagram).
The specificity of an antibody is its ability to discriminate between two different epitopes. Affinity maturation reflects an adaptive response to antigen exposure. As time goes by, the antibodies produced are able to bind more tightly to the antigen. The Capra Science Immunization Program™ allows this immunological process to fully take place. |
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Time Allows Isotype Switching
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High IgM levels are present in antiserum samples in “express”, or short immunization programs. This is not desirable and can result in confusing results in experiments for the end user.
The Capra Science Immunization Program™ allows the complete isotype switching from IgM to IgG antibodies to take place, and thus ensures antiserum with high IgG-content. |
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IMMUNIZATION TECHNIQUES
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There are
different ways of injecting the antigen:
Intradermal (ID) - Injection into the dermal layer, close to the surface Subcutaneous (SC) - Injection under epidermis in the natural cavity between the muscles and the skin. Intramuscular (IM) - Injection within the muscular tissue. Intradermal injections in multiple sites are in many antibody production facilities still often used. These injections are painful and have high risks of infection, which should in all circumstances be avoided. The skin can easily get ruptured upon the immune cell recruitment. As a result, bacteria will enter the wound and antibodies towards the infected bacteria will be generated in parallel to antigen-specific antibodies. This results in 1) a sick animal fighting a real infection and 2) a lower-titer specific antiserum containing high levels of anti-bacterial antibodies. Capra Science does not use intradermal injections for antibody production. Capra Science only uses the least painful injection technique - subcutaneous injection. Under the animal skin there is a natural cavity were the adipos/fatty layer is situated. This technique minimizes the risk for infections and the production of non-antigen specific antibodies. |
We do not use intradermal injections in Capra Science Immunization programs
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BLEEDING FREQUENCY
You may be impressed by immunization programs that promise large volumes of antiserum in relation to the animal's size. However, if the bleeds are done too frequent or in disproportional volumes to body size, both animal health and product quality are negatively affected. To compensate for frequent and large bleed volumes, the animal will 1) consume considerable amounts of liquid between bleeds and 2) put all its energy on regaining the balance of essential molecules and cells in the body, instead of "fighting" your antigen by producing high levels of specific antibody. As a consequence, your antiserum will be a diluted product with poor titer.
The Capra Science Immunization Program is formed to allow the animals to recover completely from each bleed. Our bleeds are taken at least 3-4 weeks apart. We follow the European Guidelines, where our ethical approval allows us to withdraw maximum 8ml/kg. Approximately half of the total blood volume is antiserum.
Keeping focus on the animal's health promotes generation of high-quality antiserum.
The Capra Science Immunization Program is formed to allow the animals to recover completely from each bleed. Our bleeds are taken at least 3-4 weeks apart. We follow the European Guidelines, where our ethical approval allows us to withdraw maximum 8ml/kg. Approximately half of the total blood volume is antiserum.
Keeping focus on the animal's health promotes generation of high-quality antiserum.
