CLONING AND CHARACTERIZATION OF CDNAS ENCODING V-H AND V-L OF A MONOCLONAL ANTI-CEA ANTIBODY (CEA-79) CROSS-REACTIVE WITH NCA-95 AND GENERATION OF A SINGLE-CHAIN FV MOLECULE (SCFV)
Jh. Chung et al., CLONING AND CHARACTERIZATION OF CDNAS ENCODING V-H AND V-L OF A MONOCLONAL ANTI-CEA ANTIBODY (CEA-79) CROSS-REACTIVE WITH NCA-95 AND GENERATION OF A SINGLE-CHAIN FV MOLECULE (SCFV), Molecules and cells, 7(6), 1997, pp. 816-819
We cloned complementary DNA (cDNA) encoding the variable regions of he
avy chain (V-H) and of light chain (V-L,) of a monoclonal anti-carcino
embryonic antigen (CEA) antibody cross-reactive with nonspecific cross
-reacting antigen-95 (NCA-95), which had been previously prepared and
designated as CEA 79 (gamma(2a),,, kappa). From these cDNAs, a phagemi
d expression vector for the CEA79 single chain variable fragment (scFv
) was generated. Enzyme-linked immunosorbent assay (ELISA), competitiv
e ELISAs, and Western blotting confirmed that the scFv displayed on th
e surface of the bacteriophage had retained affinity for CEA and NCA-9
5. We then determined the nucleotide sequences of the cloned cDNAs for
V-H, and V-L,. The sequence analysis revealed that V-H, and V-L, of t
he CEA 79 antibody represent new members of the mouse heavy chain subg
roup ''miscellaneous'' and the kappa light chain subgroup ''V'', respe
ctively.