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Chapter 8,335

Cloning in a bacteriophage lambda vector for the display of binding proteins on filamentous phage

Hogrefe, H.H.; Amberg, J.R.; Hay, B.N.; Sorge, J.A.; Shopes, B.

Gene 137(1): 85-91

1993

ISSN/ISBN: 0378-1119
PMID: 8282204
DOI: 10.1016/0378-1119(93)90255-2
Accession: 008334171
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We have combined the efficiency and ease of use of bacteriophage lambda vectors with the power of phage display screening technology to create SurfZAP. The use of bacteriophage lambda allows the construction of large lambda expression libraries, which are rapidly and efficiently converted to stable plasmid libraries by mass excision. In SurfZAP, clones are expressed as fusions with amino acids 198-406 of the M13 minor coat protein (cpIII) and are displayed on the surface of filamentous phage. When produced with helper phage proteins, the fusion proteins are incorporated into the surface of phagemid particles. We demonstrate the utility of biopanning by isolating tetanus toxoid-binding mouse Fab clones from SurfZAP libraries. Approximately 10-100-fold enrichment of specific clones was observed after each panning round. The ability to create a large library of genotypes and screen the phenotypes by activity may be a potent methodology for basic research and drug discovery.

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Related References

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