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Development and application of thermo‐sensitive immunomicrospheres for antibody purification
Author(s) -
Kondo Akihiko,
Kaneko Tetsuya,
Higashitani Ko
Publication year - 1994
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260440102
Subject(s) - flocculation , glycidyl methacrylate , styrene , emulsion polymerization , methacrylic acid , bovine serum albumin , chemistry , lower critical solution temperature , poly(methacrylic acid) , polymer chemistry , chromatography , polymerization , chemical engineering , copolymer , polymer , organic chemistry , engineering
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.