Cloning and sequencing of the variable region’s gene fragments of the monoclonal antibody LC 1

The whole process of cloning and sequencing of the variable regions of mAb LC 1(V H and V L) had been described. The variable regions gene fragments V H and V L of mAb LC 1 were amplified and cloned by RT PCR. The sequences of the gene fragments were determined and analyzed. The results showed that PCR product of V H region consisted of 359 bp encoding about 119 amino acid residues, and PCR product of V L region contained 337 bp encoding about 112 amino acid residues. The amino acid sequences of polypeptide V H and V L were in agreement with the characterization of the amino acid sequences of known mouse antibodies’ variable region. According to Kabat classified method, mAb LC 1 V H gene segment and V L gene segment belong to the mouse Ig heavy chain subgroup V and κ chain subgroup V κ 21 respectively. The success of cloning of the gene fragments of mAb LC 1’s variable regions lay a benefit basis for the construction and expression of single chain Fv or Fab.