A method for linking VL and VH region genes that allows bulk transfer between vectors for use in generating polyclonal IgG libraries.

Published

Journal Article

Libraries of Ab fragments have been produced by others from light and heavy chain cDNAs derived from populations of B lymphocytes and expressed in bacteria. However, such libraries have not been transferred to eukaryotic expression vectors to generate polyclonal libraries of intact glycosylated Abs that can mediate effector functions. We present a method for transferring pairs of linked VL-VH region genes between circular prokaryotic and eukaryotic vectors. The key feature of the transfer is that the VL and VH region genes are linked head to head (<-->) in opposite transcriptional orientations. To illustrate this method, a pair of VL and VH region cDNAs derived from an existing hybridoma cell line were linked head to head by PCR, transferred as a unit between vectors, and expressed as an IgG Ab with Ag binding activity. Although we tested the transfer of a single VL-VH region gene pair, this system is expected to allow the bulk transfer of physically linked VL-VH region gene combinations between different circular vectors and the expression of the same library as either Ab fragments or intact Abs.

Full Text

Duke Authors

Cited Authors

  • Sarantopoulos, S; Kao, CY; Den, W; Sharon, J

Published Date

  • June 1, 1994

Published In

Volume / Issue

  • 152 / 11

Start / End Page

  • 5344 - 5351

PubMed ID

  • 8189052

Pubmed Central ID

  • 8189052

Electronic International Standard Serial Number (EISSN)

  • 1550-6606

International Standard Serial Number (ISSN)

  • 0022-1767

Language

  • eng