Cloning of cDNAs coding for heavy- and light-chain variable regions of a murine monoclonal antibody with specificity for human plasma apolipoprotein A-I.
We prepared a monoclonal antibody (mAb) A-I4-48 to HDL apolipoprotein A-I with the ultimate goals of expressing the valuable immunodiagnostic single-chain Fv (scFv) in Escherichia coli. The binding specificity of mAb A-I4-48 was determined by ELISA and Western blot analysis. The dissociation constant (Kd) of antigen-antibody complex obtained by ELISA was 8.33 x 10(-8) M. Poly(A)+ RNA was prepared from the hybridoma cell line secreting mAb A-I4-48 and used as a template for cDNA synthesis and cloning. The nucleotide sequence analyses revealed that the variable regions of the heavy- and light-chains were members of mouse heavy-chain subgroup IIA and kappa light-chain subgroup II, respectively. Comparison of the nucleotide sequences with mouse immunoglobulin genes in the GenBank data base showed that the cDNAs have not been previously reported.
Lee
S H
SH
Department of Biochemistry, College of Medicine, Soonchunhyang University, Choong-Nam, South Korea.
Kim
C S
CS
Cho
M H
MH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D016632:Apolipoprotein A-I / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D016632:Apolipoprotein A-I; D018076:DNA, Complementary; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region