Mol. Cells 1997; 7(6): 816~819  
Cloning and Characterization of cDNAs encoding theVH and VL of a Monoclonal Anti-CEA Antibody (CEA 79) Cross-Reactive with NCA-95 and Generation of a Single-chain Fv Molecule (scFv)
Jun Ho Chung, Suk Jin Choi Hyo Jung Gm Ik Jung Gm In Hak Choi, Sang
Do Lee Kye Sook Yi Pann Ghill Suh Sung Ho Ryu and Hong Keun Chung
; Published online December 31, 1997.
© The Korean Society for Molecular and Cellular Biology. All rights reserved.

ABSTRACT
We cloned complementary DNA (cDNA) encoding the variable regions of heavy chain (VH) and of light chain (VL) of a monoclonal anti-carcinoembryonic antigen (CEA) antibody cross-reactive with nonspecific cross-reacting antigen-95 (NCA-95), which had been previously prepared and designated as CEA 79 (γ2a, κ). From these cDNAs, a phagemid expression vector for the CEA79 single chain variable fragment (scFv) was generated. Enzyme-linked immunosorbent assay (ELISA), competitive ELISAs, and Western blotting confirmed that the scFv displayed on the surface of the bacteriophage had retained affinity for CEA and NCA-95. We then determined the nucleotide sequences of the cloned cDNAs for VH and VL . The sequence analysis revealed that VH and VL of the CEA 79 antibody represent new members of the mouse heavy chain subgroup "miscellaneous" and the kappa light chain subgroup "V", respectively.

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