Abstract
Immunoglobulin heavy chain Fd gene and κ chain gene were amplified by Polymerase Chain Reaction (PCR) from total RNA of a hybridoma cell line secreting anti-stomach cancer monoclonal antibody 3G9. Sequences analysis showed that VH, D, Jhgenes were derived from VH7183, DFL16. 1, and JH3, and that Vk and Jk from VkII and J.2.3G9 Fd and κ DNA segments were cloned into expression vector pComb3. Soluble Fab was expressed by IPTG induction and proved by western blotting. The specific antigen binding activity of the bacterially produced 3G9 Fab was demonstrated by enzyme- linked immunofiltration assay.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
1992, 53
Better M, et al. Escherichia coli secretion of an active chimeric antibody fragment. Science 1988; 240: 1041.
Barbas III CF, et al. Assembly of combinatorial antibody libraries on phage surfase: the gene III site. Pro Natl Acad Sci USA 1991; 88: 7978.
1989;11(3):162.
Chomozynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156.
Huse WD, et al. Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science 1989; 246: 1275.
1985;5(1):33d.
Kabat EA, et al. Sequences of proteins of immunological interest. 5th Edition. Bethesda: US. Department of Health and Human Service. 1991.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Wang, Y., Xu, J., Li, Y. et al. Cloning and expression of Fab genes of anti-stomach cancer McAb 3G9 inE. coli . Chin J Cancer Res 5, 1–6 (1993). https://doi.org/10.1007/BF02997485
Issue Date:
DOI: https://doi.org/10.1007/BF02997485