Immunoglobulin Applications

Immunoglobulins have been used in applications from diagnostic assays to therapeutics. The most widely used immunoglobulin, IgG, is derived from mammals. Chickens, however, produce a slightly different antibody, IgY, which contains a longer Fc region.

What makes chickens an attractive source of antibody production is the greater evolutionary distance between chickens and mammals. Chickens, for the most part, do not recognize mammalian proteins as “self”. These “self” proteins become more immunogenic, leading to an increased antibody diversity in chickens when compared to the mammalian host.

There are benefits of using IgY for prophylactic and therapeutic non-systemic applications. IgY does not activate the mammalian complement, react with rheumatoid factors, or bind to mammalian Fc receptors. Due to the stability of IgY at low pH conditions, it can be used to combat gastrointestinal infections.

We have been involved in the development of gastrointestinal treatments using IgY in agriculture as well as respiratory treatments to prevent COVID-19.

Why Use Specific-Pathogen-Free (SPF) Hens for Your IgY Antibody Production?

Our hens are kept in filtered-air, positive-pressure rooms with strict biosecurity. Hens under these conditions have a naïve immune system, which allows for them to ramp up IgY production quickly compared to commercial layers. SPF hens are also free of pathogens as listed under United States Department of Agriculture (USDA) and European Pharmacopeia (Ph. Eur.) guidelines leading to less risk of contamination in your product.

Chicken IgY Antibody Production Process

Our standard chicken IgY antibody production process includes three SPF hens per customer supplied antigen. One hundred eggs are collected over a three-month period. Pre-immune blood samples are tested on ELISA and Western Blot before each bird is inoculated with antigen. Boosts and serum sampling occur every other week until week six at which point egg collection begins. Boosts and serum sampling continue every two weeks, and the yolks are separated from the eggs and frozen on a weekly basis until 100 yolks in total are processed. IgY antibody extraction occurs via water dilution and pH change and is concentrated and bottled for shipment on dry ice.

Chicken Immunization Strategy

poster of avian inoculation with antigen

Standard 100 Egg Yolk IgY Project

  • The following elements are involved in our standard IgY antibodies project
    Specifications Included
    Protein name (for label and CofA wording)   
    Number of SPF chickens (starting number; the number of hens laying after inoculation may be less)  3
    Number of eggs to collect  100
    Quantity of protein needed from client  18 mg @ >/= 2 mg/ml
    Quantity of protein per inoculation per bird  500 µg 
    Inoculum volume per bird (0.5 ml max)  0.5 ml 
    Two-day bioburden test on protein  NA - with sterility results or sterile filtration 
    Pre-immune bleed - tested via ELISA and Western blot  Y
    Initial inoculation with Freund's Adjuvant  Y
    Boost 1 at 2 weeks with Incomplete Freund's Adjuvant (ICF)  Y
    Serum sample at 2 weeks - Tested via ELISA and Western blot  Y
    Boost 2 at 4 weeks with ICF  Y
    Serum sample at 4 weeks - Tested via ELISA and Western blot  Y
    Boost 3 at 6 weeks with ICF  Y
    Egg collection starts at 6 weeks  Y
    Serum sample at 6 weeks - Tested via ELISA and Western blot  Y
    Boost 4 @ 8 weeks with ICF  Y
    Serum sample at 8 weeks - Tested via ELISA & Western blot  Y
    Boost 5 at 10 weeks with ICF  Y
    Serum sample at 10 weeks - Tested via ELISA & Western blot  Y
    Termination at 12 weeks or optional extension  Y
    IgY antibody extraction from egg yolks via water dilution and pH change  Y
    Bottled in 45 ml aliquots using conical tubes  Y
    Final chicken IgY antibodies tested via ELISA and Western blot  Y

     

Technician performing assay for host cell proteins (HCPs) in a lab.

Host Cell Proteins: New Tools for an Ongoing Challenge

This on-demand webinar delves into testing regulations for host cell proteins (HCPs), the potential risks of using inappropriate methods, and different approaches you can take to protect your biopharmaceuticals.

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3R Impact

SPF chickens are efficient producers of immunoglobulin Y antibodies and a single egg yolk can generate 100-200 mg of IgY; to put it into perspective, one hen can produce in one months' time the equivalent quantity of antibodies in their eggs as a goat. This fits perfectly into the principles of the 3Rs (Replacement, Reduction, and Refinement).