Human κ chain expression in a λ phage vector: methods of isolating amplified cDNA affects cloning efficiency

Article type: Research Article

Authors: Lazaridis, Konstantinos | Dang, Howard^; | Talal, Norman

Affiliations: Audie L. Murphy Memorial Veterans Hospital and the University of Texas Health Science Center, San Antonio, TX, USA

Note: [] Address reprint requests to Dr. Howard Dang, Department of Medicine, Division of Clinical Immunology, 7703 Floyd Curl Drive, San Antonio, TX 78284-7874, USA.

Abstract: Three common methods of isolating amplified DNA were evaluated for their effects on cloning efficiency in a phage vector designed to express human κ chain. The “glass milk” technique gave higher cloning efficiency and protein expression than phenol-chloroform extraction or microfiltration. This shows that the quality of amplified cDNA should be considered when studying the human antibody repertoire using this vector system.

Keywords: recombinant antibody, polymerase chain reaction, DNA isolation

DOI: 10.3233/HAB-1993-4302

Journal: Human Antibodies, vol. 4, no. 3, pp. 104-106, 1993