Cross-linked filamentous phage as an affinity matrix.
Filamentous phage can be cross-linked to make a hydrophilic aggregate that is pelleted by low-speed centrifugation. The aggregate is stable at near-neutral pHs, and withstands exposure to the acid buffers (pH down to 2.2) that are often used as eluents in immunoaffinity purification. If a peptide epitope is genetically fused to a coat protein on the virion surface, the aggregate serves as an effective affinity matrix for absorbing and affinity-purifying antibodies that bind the peptide. When the peptide epitope is first obtained in this form by selection from large phage display libraries, this ability to fashion an affinity matrix directly from the selected phage represents a significant streamlining of research and development.
Smith
G P
GP
Division of Biological Sciences, University of Missouri, Columbia 65211, USA. gpsmith@biosci.mbp.missouri.edu
Petrenko
V A
VA
Matthews
L J
LJ
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000302:isolation & purification; D000918:Antibody Specificity; D000956:Antigens, Viral; D017122:Bacteriophage T4 / Q000276:immunology; D002213:Capsid / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D003432:Cross-Linking Reagents; D004279:DNA, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000302:isolation & purification; D017912:Inovirus / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D011817:Rabbits; D014771:Virion / Q000276:immunology / Q000378:metabolism
D000914:Antibodies, Viral; D000956:Antigens, Viral; D003432:Cross-Linking Reagents; D004279:DNA, Viral; D000939:Epitopes