Controlled growth of Chinese hamster ovary cells and high expression of antibody-IL-2 fusion proteins by temperature manipulation.
Growth and the expression of the anti-ErbB2 scFv-Fc-IL2 fusion protein in Chinese hamster ovary (CHO) cells were in association at 37 degrees C. The expression of the fusion protein was no more than 25 microg/ml. At 30 degrees C the cell growth was arrested but the cells continued to produce the fusion protein up to 60-80 microg/ml. About 50% of CHO cells were rapidly blocked in G2/M phase after the temperature was shifted from 37 to 30 degrees C. Lowering temperature resulted in cell growth arrest, but maintained cell viability for a longer time and enhanced the production of the antibody-IL-2 fusion protein in CHO cells.
Shi
Ming
M
Department of Molecular Immunology, Institute of Basic Medical Sciences, 27 Taiping Road, 100850 Beijing, P.R. China.
Xie
Zhigang
Z
Yu
Ming
M
Shen
Beifen
B
Guo
Ning
N
D000818:Animals; D000906:Antibodies / Q000235:genetics*; D017209:Apoptosis / Q000502:physiology; D016466:CHO Cells; D002453:Cell Cycle / Q000502:physiology; D049109:Cell Proliferation; D002470:Cell Survival / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005784:Gene Amplification / Q000187:drug effects / Q000235:genetics; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000235:genetics; D007074:Immunoglobulin G / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics*; D008727:Methotrexate / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013696:Temperature; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection
D000906:Antibodies; D007141:Immunoglobulin Fc Fragments; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D007376:Interleukin-2; D008727:Methotrexate; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; D013762:Tetrahydrofolate Dehydrogenase