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 language | English |
|---|---|
| Pages (from-to) | 570-580 |
| Number of pages | 11 |
| Journal | Biotechnology and Bioengineering |
| Volume | 75 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - Dec 5 2001 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
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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 journal › Article
}
TY - JOUR
T1 - Type-specific separation of animal cells in aqueous two-phase systems using antibody conjugates with temperature-sensitive polymers
AU - Kumar, A.
AU - Kamihira, M.
AU - Galaev, I. Yu
AU - Mattiasson, B.
AU - Iijima, S.
PY - 2001/12/5
Y1 - 2001/12/5
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0035814330&partnerID=8YFLogxK
U2 - 10.1002/bit.10080
DO - 10.1002/bit.10080
M3 - Article
C2 - 11745133
AN - SCOPUS:0035814330
VL - 75
SP - 570
EP - 580
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
SN - 0006-3592
IS - 5
ER -