Cloning and characterization of cDNAs encoding VH and VL of a monoclonal anti-CEA antibody (CEA 79) cross-reactive with NCA-95 and generation of a single-chain Fv molecule (scFv).
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 (gamma 2a, kappa). 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.
Chung
J H
JH
Department of Biochemistry, College of Medicine, Dongguk University, Kyungju, Korea.
Choi
S J
SJ
Kim
H J
HJ
Kim
I J
IJ
Choi
I H
IH
Lee
S D
SD
Yi
K S
KS
Suh
P G
PG
Ryu
S H
SH
Chung
H K
HK
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000951:Antigens, Neoplasm; D001435:Bacteriophages / Q000276:immunology; D001483:Base Sequence; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D015815:Cell Adhesion Molecules; D003001:Cloning, Molecular; D003429:Cross Reactions / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; C111832:CEA 79 scFv; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D008562:Membrane Glycoproteins; D057127:Single-Chain Antibodies