Type-specific separation of animal cells in aqueous two-phase systems using antibody conjugates with temperature-sensitive polymers

A. Kumar, M. Kamihira, I. Yu Galaev, B. Mattiasson, S. Iijima

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

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-l) 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.

Original languageEnglish
Pages (from-to)570-580
Number of pages11
JournalBiotechnology and Bioengineering
Volume75
Issue number5
DOIs
Publication statusPublished - Dec 5 2001
Externally publishedYes

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Cell Separation
Antibodies
Polymers
Animals
Cells
Temperature
Ligands
Monoclonal antibodies
Dextran
Anhydrides
T-Cell Lymphoma
Myeloid Cells
Dextrans
Acute Myeloid Leukemia
Polyethylene glycols
Monoclonal Antibodies
poly-N-isopropylacrylamide

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Type-specific separation of animal cells in aqueous two-phase systems using antibody conjugates with temperature-sensitive polymers. / Kumar, A.; Kamihira, M.; Galaev, I. Yu; Mattiasson, B.; Iijima, S.

In: Biotechnology and Bioengineering, Vol. 75, No. 5, 05.12.2001, p. 570-580.

Research output: Contribution to journalArticle

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