| Comments |
This line is Neomycin resistant.
The cell line was derived by co-transfecting the CHO cell line DXB11 using Lipofection with heavy and light chain expression plasmids constructed from the antibody produced by the 23F2G cell line (ATCC-HB-11081).
The plasmids contain both cytomegalovirus (CMV) and SV40 viral sequences.
The 23F2G antibody is reactive with the beta chain (CD18) of human leukocyte integrins.
The cell line is capable of producing about 100 micrograms/liter of the humanized immunoglobulin at the rate of 25 pg/cell/day. |
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| References |
Rose LM. Murine and humanizer 23F2G antibodies and cell lines expressing said antibodies. US Patent 5,854,070 dated Dec 29 1998
Bowen JD, et al. Phase I study of a humanized anti-CD11/CD18 monoclonal antibody in
multiple sclerosis. Clin. Pharmacol. Ther. 64: 339-346, 1998. PubMed: 9757158
Yenari MA, et al. Hu23F2G, an antibody recognizing the leukocyte CD11/CD18 integrin,
reduces injury in a rabbit model of transient focal cerebral ischemia. Exp. Neurol. 153: 223-233, 1998. PubMed: 9784282
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
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