Isolation of alpha-crystallin and its subunits by affinity chromatography on immobilized monoclonal antibodies.
Use has been made of the specific interactions between monoclonal antibodies and the alpha A or alpha B subunits of alpha-crystallin to devise methods for the purification of the intact protein or its subunits. alpha A and alpha B subunits were separated by affinity chromatography on an immobilized monoclonal antibody specific for alpha A chains, using a pH gradient. Use of an antibody which binds both subunits has enabled the isolation of intact alpha-crystallin aggregates. Gel electrophoresis, conformational probing and size analysis showed that the affinity purified proteins were purer but otherwise indistinguishable from the alpha-crystallins isolated by other methods.
Stevens
A
A
Russell Grimwade School of Biochemistry, University of Melbourne, Parkville, Victoria, Australia.
Augusteyn
R C
RC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002417:Cattle; D002846:Chromatography, Affinity; D003459:Crystallins / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D011485:Protein Binding; D011487:Protein Conformation
D000911:Antibodies, Monoclonal; D003459:Crystallins