Development and application of thermo‐sensitive immunomicrospheres for antibody purification

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Abstract
The latex particles composed of poly(styrene/N‐isopropylacrylamide/glycidyl methacrylate) [P(St/NIPAM/GMA)] and poly(styrene/N‐isopropylacrylamide/methacrylic acid) [P(St/NIPAM/MAA)] were prepared by emulsifier‐free emulsion polymerization. These latex particles with submicrometer size showed the thermosensitivity originated from the thermo‐sensitive nature of NIPAM. That is, the minimum NaCI concentration for flocculation of these latex particles [critical flocculation concentration (CFC)] decreased significantly with increasing temperature and reached constant values at above the critical temperature [critical flocculation temperature (CFT)]. At a certain NaCl concentration, the thermo‐sensitive latex particles were flocculated by raising temperature, and conversely, the flocculated thermo‐sensitive latex particles were completely dispersed by lowering temperature. Bovine serum albumin (BSA) was covalently immobilized onto the P(St/NIPAM/GMA) and P(St/NIPAM/MMA) latex particles with high efficiency. The BSA‐immobilized P(St/NIPAM/GMA) and P(St/NIPAM/MAA) latex particles (immunomicrospheres) showed the similar dependencies of CFC on temperature to the bare latex particles. These thermo‐sensitive immunomicrospheres were successfully used for the immunoaffinity purification of anti‐BSA antibodies from antiserum. © 1994 John Wiley & Sons, Inc.