A Refined Approach to Producing Polyclonal Antibodies in Chickens – Completely Replacing All Invasive Elements by Combining Immunizations with Routine Aerosol-based Vaccinations

Contact: Otto Kalliokoski

The aim is to exploit routine vaccination of chickens, administered in aerosol form, to raise antibodies against a model immunogen (a protein) coupled to the surface of the vaccine’s virus particles. Combined with isolation/purification of antibodies from egg yolk, this method will not only reduce the number of animals needed in the production of polyclonal antibodies, but also completely eliminate the need of invasive procedures associated with conventional antibody production, thus constituting an immense refinement of methodology.

Project status at January 2017

The goal of our project is to immunize chickens for antibody production without ever having to touch or restrain them. By attaching the immunogen (the antibody target) onto the virus particles of an inhalable vaccine which all chickens in Denmark is given after birth, we believe that we can provide immunity to the disease (infectious bronchitis) while simultaneously producing antibodies. We have previously developed methods for extracting antibodies from hens’ eggs and within the project we have refined our methods for detecting immunity toward the vaccine based on the hen’s antibodies. In addition we have confirmed that we can successfully immunize chickens with quantities of immunogen small enough to be carried on the surface of virus particles and developed a method for combining the two (and methods for evaluating the conjugation process). Finally, we have developed a method for delivering a microliter-scale aerosol to day-old chickens without having to restrain them.

Looking back on the previous year, our project has come a long way, but one unforeseen problem has prevented us from putting all the elements of the project together for a final test. Chicken vaccines are not developed to be nearly as pure as humane vaccines. Using electron microscopy we found that the Commercial vaccines contained more – by several orders of magnitude – impurities and stabilizing ingredients than they contained actual attenuated virus. These impurities interfere with the conjugation methods we have developed, effectively preventing us from attaching the immunogen to the virus. We are therefore currently working on a method for of purifying the virus particles from the commercial vaccine preparation in a gentle enough manner to not damage its virulence. We are currently holding off on carrying out a final experiment on chickens before we believe that we have a satisfactory purity of the starting material and can test our hypothesis under optimal conditions.

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