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Type‐specific separation of animal cells in aqueous two‐phase systems using antibody conjugates with temperature‐sensitive polymers
Author(s) -
Kumar A.,
Kamihira M.,
Galaev I. Yu.,
Mattiasson B.,
Iijima S.
Publication year - 2001
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.10080
Subject(s) - conjugate , chemistry , aqueous solution , polymer , lower critical solution temperature , dextran , polyethylene glycol , conjugated system , polymer chemistry , chromatography , organic chemistry , copolymer , mathematical analysis , mathematics
A new type of aqueous two‐phase system (ATPS) has been developed in which a temperature‐sensitive polymer, poly‐ N ‐isopropylacrylamide [poly (NIPAM)] was used as a ligand carrier for the specific separation of animal cells. Monoclonal antibodies were modified with itaconic anhydride and copolymerized with N ‐isopropylacrylamide, and the ligand‐conjugated carriers were added to the polyethylene glycol 8000–dextran T500 aqueous two‐phase systems. The antibody–polymer conjugates were partitioned to the top phase in the absence or presence of 0.15 M NaCl. When ligand‐conjugated carriers were used, more than 80% of the cells were specifically partitioned to the top phase in the presence of NaCl up to 0.1 M. The cells were partitioned almost completely to the bottom phase at 0.1 M NaCl or above, when no antibody–conjugate was added in the ATPS. As a model system, CD34‐positive human acute myeloid leukemia cells (KG‐1) were specifically separated from human T lymphoma cells (Jurkat) by applying anti‐CD34 conjugated with poly‐ N ‐isopropylacrylamide in the aqueous two‐phase system. By the temperature‐induced precipitation of the polymer, about 90% of the antibody–polymer conjugates were recovered from the top phase, which gave approximately 75% cell separating efficiency in the next cycle of reuse. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 570–580, 2001.