Chicken is an excellent choice of host, and the phylogenetic distance to mammals make chicken-antibodies suitable for most projects. What is more, for anti-peptide antibody projects, where we have designed and produced the peptide, we will guarantee that the resulting IgY will contain antibodies that target the peptide.
Of course, you can fully customise your project, but for your convenience we have outlined a typical project.
An immunogen is required for the antibody production. This can be a protein or other molecule that you provide, or it can be produced here. In the latter case, either the entire protein can be obtained using a recombinant technique, or a peptide is designed and synthesized. Using peptides as antigens is the most frequently used strategy. It should be noted that the more effort that is put into this step, the better the odds for the successful outcome of your project. Other labs may choose to ignore this fact, but the production of high quality peptides for antibody production may very well be pivotal for the success of your project.
If you will be supplying the immunogen, there are a few things to keep in mind;
One or two chickens are immunised using our 11 week standard protocol. For each chicken one to two pre-immune eggs are collected prior to immunisations, and ten immune eggs are collected towards the end of the immunisation schedule. Each egg yolk contains approximately 100 mg total IgY that can be extracted. Thus, chickens produce large amounts of antibody at a rather low cost per unit.
Managing your project will ensure that you obtain antibody in the required amounts, while keeping costs down. This means collecting a sufficient number of eggs, isolating the IgY, and then terminating the project as soon as possible.
The egg yolk contains the IgY, which needs to be extracted before use. We will isolate the IgY fraction from the egg yolk of the pre-immune eggs and four immune eggs. The IgY is then tested by ELISA. The typical yield is about 400 mg of total IgY from the immune eggs. The remaining six immune eggs are stored for a while in order for you to evaluate the first batch.
The IgY can be purified further using an antigen specific affinity column in order to obtain the specific antibodies. The expected yield is a few per-cent of the total IgY.
Examples of references of customers using our custom chicken antibody services include;
Distribution of neuronal KCC2a and KCC2b isoforms in mouse CNS
M. Markkanen, T. Karhunen, O. Llano, A. Ludwig, C. Rivera, P. Uvarov and M. S. Airaksinen
Journal of Comparative Neurology, Volume 522, Issue 8, pages 1897–1914, 01 June 2014
Irgm1 (LRG-47), a Regulator of Cell-Autonomous Immunity, Does Not Localize to Mycobacterial or Listerial Phagosomes in IFN-γ–Induced Mouse Cells
H. M. Springer, M. Schramm, G. A. Taylor and J. C. Howard
The Journal of Immunology August 15, 2013 vol. 191 no. 4 1765-1774
A unique cell division machinery in the Archaea
A.-C. Lindås, E. A. Karlsson, M. T. Lindgren, T. J. G. Ettema, and R. Bernander. 2008.
PNAS December 2, 2008 vol. 105 no. 48 18942-18946.
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