Type-specific separation of animal cells in aqueous two-phase systems using antibody conjugates with temperature-sensitive polymers.
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.
Kumar
A
A
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya 464-8603, Japan.
Kamihira
M
M
Galaev
I Y
IY
Mattiasson
B
B
Iijima
S
S
D000178:Acrylamides / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D018952:Antigens, CD34 / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D017743:Equipment Reuse; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D019169:Jurkat Cells; D007951:Leukemia, Myeloid; D011108:Polymers / Q000737:chemistry; D013696:Temperature; D014407:Tumor Cells, Cultured
D000178:Acrylamides; D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D011108:Polymers; C067295:N-isopropylacrylamide