title: Purification of Antibodies Using Affinity Chromatography.
abstract: Affinity chromatography permits the isolation of a target analyte from a complex mixture and can be utilized to purify proteins, carbohydrates, drugs, haptens, or any analyte of interest once an affinity pair is available. It involves the exploitation of specific interactions between a binding affinity pair, such as those between an antibody and its associated antigen, or between any ligand and its associated binding receptor/protein. With the discovery of protein A in 1970, and, subsequently protein G and L, immuno-affinity chromatography has grown in popularity and is now the standard methodology for the purification of antibodies which may be implemented for a selection of different applications such as immunodiagnostics. This chapter is designed to inform the researcher about the basic techniques involved in the affinity chromatography-based purification of monoclonal, polyclonal, and recombinant antibodies. Examples are provided for the use of protein A and G. In addition, tables are provided that allow the reader to select the most appropriate protein for use in the isolation of their antibody.
authors.affiliation: School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.
authors.forename: Elaine
authors.initials: E
authors.lastname: Darcy
authors.affiliation: School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.|Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland.
authors.forename: Paul
authors.initials: P
authors.lastname: Leonard
authors.affiliation: School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland.
authors.forename: Jenny
authors.initials: J
authors.lastname: Fitzgerald
authors.affiliation: Ashtown Food Research Centre, Teagasc, Ashtown, Dublin 15, Ireland.
authors.forename: Martin
authors.initials: M
authors.lastname: Danaher
authors.affiliation: Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland.
authors.forename: Hui
authors.initials: H
authors.lastname: Ma
authors.affiliation: School of Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland. Richard.okennedy@dcu.ie.|Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland. Richard.okennedy@dcu.ie.
authors.forename: Richard
authors.initials: R
authors.lastname: O'Kennedy
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry; D011232:Chemical Precipitation; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; C055047:Ig L-binding protein, Peptostreptococcus; C034284:IgG Fc-binding protein, Streptococcus; D013205:Staphylococcal Protein A
keywords: Affinity chromatography; Monoclonal; Polyclonal; Protein A; Protein G; Protein L; Recombinant antibodies