Target-selective homologous recombination cloning for high-throughput generation of monoclonal antibodies from single plasma cells.

ABSTRACT:

Background

Molecular cloning of functional immunoglobulin genes from single plasma cells is one of the most promising technologies for the rapid development of monoclonal antibody drugs. However, the proper insertion of PCR-amplified immunoglobulin genes into expression vectors remains an obstacle to the high-throughput production of recombinant monoclonal antibodies.

Results

We developed a single-step cloning method, target-selective homologous recombination (TS-HR), in which PCR-amplified immunoglobulin variable genes were selectively inserted into vectors, even in the presence of nonspecifically amplified DNA. TS-HR utilizes Red/ET-mediated homologous recombination with a target-selective vector (TS-vector) with unique homology arms on its termini. Using TS-HR, immunoglob

SUBMITTER: Kurosawa N 

PROVIDER: S-EPMC3088891 | biostudies-literature | 2011 Apr

REPOSITORIES: biostudies-literature