[Amplification and sequencing of heavy chain variable region (VH) gene of a human monoclonal antibody (CM-1) with specificity to breast cancer].
For rescueing the heavy chain variable region (VH) gene of CM-1--a human monoclonal antibody against breast cancer, total RNA was extracted from the CM-1 hybridoma cells. After preparing its cDNA by reverse transcription and amplifying by PCR, a DNA fragment with about 400 bp was obtained. With an internal primer, the fragment was directly sequenced and 344 bases which consisted well to known human antibody VH gene construction were read out. In addition, its corresponding amino acid sequences coincided with the 1-3 group canonical structure of human antibody variable region. So we suggested that CM-1 could be considered as an individual of human antibody family, and should aid the preparation of antibody molecules.
Chen
L
L
Institute of Basic Medical Sciences, CAMS, Beijing.
Wang
S
S
Zhang
Y
Y
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics*; D001483:Base Sequence; D001943:Breast Neoplasms / Q000276:immunology*; D018076:DNA, Complementary / Q000235:genetics; D005784:Gene Amplification; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007075:Immunoglobulin M / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007143:Immunoglobulin Heavy Chains; D007075:Immunoglobulin M; D007135:Immunoglobulin Variable Region