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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Produce Safety and Microbiology Research » Research » Publications at this Location » Publication #305636

Research Project: Immunodiagnostics to Detect Prions and Other Important Animal Pathogens

Location: Produce Safety and Microbiology Research

Title: Affinity purification of antibodies

Author
item Hnasko, Robert
item McGarvey, Jeffery - Jeff

Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: 4/4/2015
Publication Date: 7/14/2015
Citation: Hnasko, R.M., McGarvey, J.A. 2015. Affinity purification of antibodies. In: Hnasko, R., editor. ELISA Methods and Protocols. New York, NY: Humana Press. p. 29-41. doi: 10.1007/978-1-4939-2742-5.

Interpretive Summary: This Manuscript describes methods used in the purification of antibodies from different sources.

Technical Abstract: Antibodies are provided in a variety of formats that includes antiserum, hybridoma culture supernatant or ascites. They can all be used successfully in crude form for the detection of target antigens by immunoassay. However, it is advantageous to use purified antibody in defined quantity to facilitate assay reproducibility, economy, and reduced interference of non--specific components as well as improved storage, stability and bio--conjugation. Although not always necessary, the relative simplicity of antibody purification using commercially available protein--A, --G or --L resins with basic chromatographic principles warrants purification when antibody source material is available in sufficient quantity. Here we define three simple methods using immobilized 1) protein--A, 2) protein--G, and 3) protein--L agarose beads to yield highly purified antibody.