cDNA sequence analysis of monoclonal antibodies against the human placental acidic isoferritin.
By using human placental acidic isoferritin (PAF) as antigen to immunize BALB/c mice and conventional cell fusion, we have established three mouse hybridoma cell lines that secrete IgG monoclonal antibodies (MAbs) to PAF, termed as Z-2-3, Z-2-5, and Z-3-6, respectively. In ELISA, the MAbs were shown to react specifically with human PAF. We then applied the polymerase chain reaction (PCR) technique to clone variable region genes of the heavy (V(H)) and light (V(L)) chains of these MAbs, and appropriate full-length cDNA clones were obtained and characterized by nucleotide sequence analysis. V(H) and V(L) segments of anti-human PAF MAbs belong to the J558 and Vkappa19 family, respectively. The nucleotide sequence of Z-3-6 in the V(H) segment is highly homologous to that of MAb 18.1.16 except for their diversity minigenes. The light chain sequences of these MAbs show high homology with that of MAb cc92. It is implied that the D segment and the nucleotides inserted at the V(H)-D and D-J splice junctions are mostly responsible for the specificity of Z-3-6, and that the differences between the V(H) and V(L) sequences of these MAbs may determine their different affinity or recognition of different antigenic determinants.
Department of Immunology, Tongji Medical University, Wuhan, PR China.
C
C
Zhou
J
J
Shao
X
X
Wang
H
H
Zhu
Y
Y
Zhang
W
W
Xiong
G
G
Shen
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics*; D005260:Female; D005293:Ferritins / Q000276:immunology*; D005803:Genes, Immunoglobulin; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010920:Placenta / Q000276:immunology*; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D005293:Ferritins