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Nucleic Acids Research, 2000, Vol. 28, No. 20 e85
© 2000 Oxford University Press

Amplification of IgG VH and VL (Fab) from single human plasma cells and B cells

J. A. Coronella, P. Telleman, T. D. Truong, F. Ylera and R. P. Junghans*

Biotherapeutics Development Lab, Harvard Institute of Human Genetics, Harvard Medical School and Beth Israel Deaconess Medical Center, Boston, MA 02215, USA

Amplification of human immunoglobulin has many potential applications such as analysis of clonality, isolation of immunogenic antigens and antigen-specific immunotherapy. Here we describe a method for amplification of human immunoglobulin heavy and light chains from single B lymphocytes or plasma cells. Cells are isolated by FACS, and Ig is amplified by semi-nested RT–PCR. The method is versatile, sensitive and reliable: it provides appropriately paired heavy and light chains, requiring as little as 2 days to produce amplified Fab DNA from human tissues.

* To whom correspondence should be addressed at: 4 Blackfan Circle, HIM 403, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA. Tel: +1 617 432 7004; Fax: +1 617 432 7007; Email: junghans@hms.harvard.edu


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