Aim: To seek a good method for simultaneous cloning V(L), V(H) and signal peptide genes of anti-CD20 monoclonal antibody (mAb) from hybridoma cells 1-28.
Methods: Total RNA was prepared from hybridoma cells 1-28 by Trizol. Two methods were used to clone the target genes. One was traditional rapid amplification of 5' cDNA end (T-5' RACE), and the other was RNA ligase-mediated rapid amplification of 5' cDNA end (RLM-RACE). The PCR products were then cloned into pGEM-T Easy vector, sequenced and analyzed with Blast method using Kabat and GenBank Databases.
Results: The target genes of L chain and H chain were both successfully cloned by RLM-RACE, while only V(L) gene and its signal peptide genes were obtained by T-5' RACE.
Conclusion: RLM-RACE is a good method for cloning antibody's V gene and signal peptide genes.