title: Affinity Purification of Antibodies.
abstract: Antibodies are provided in a variety of formats that include 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 nonspecific 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, protein-G, or protein-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.
authors.affiliation: Produce Safety and Microbiology Unit (PSM), Western Regional Research Center (WRRC), Pacific West Area (PWA), Agricultural Research Service (ARS), United States Department of Agriculture (USDA), 800 Buchanan St., Albany, CA, 94710, USA, Robert.hnasko@ars.usda.gov.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hnasko
authors.forename: Jeffery A
authors.initials: JA
authors.lastname: McGarvey
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001201:Ascites / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D017077:Culture Media, Conditioned / Q000737:chemistry; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry; D007106:Immune Sera / Q000737:chemistry; D007132:Immunoglobulin Isotypes / Q000737:chemistry; D051379:Mice; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; C055047:Ig L-binding protein, Peptostreptococcus; C034284:IgG Fc-binding protein, Streptococcus; D055767:Immobilized Proteins; D007106:Immune Sera; D007132:Immunoglobulin Isotypes; D013205:Staphylococcal Protein A