Abstract
Expression cloning of a mouse kappa chain fragment has been achieved from a cDNA library by display of expressed proteins on filamentous phage and affinity selection for binding to anti-mouse Fab antibodies. Expressed proteins were anchored to the phage coat by a synthetic, anti-parallel leucine zipper, which had been selected from a semi-randomized zipper library for the ability to connect a test protein to phage. From a library of 4 x 10(6) transformants, two separate clones displaying different size cDNA inserts were recovered after four selection rounds. These results further demonstrate the utility of phage display for cDNA expression cloning.
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- Clackson T., Wells J. A. In vitro selection from protein and peptide libraries. Trends Biotechnol. 1994 May;12(5):173–184. doi: 10.1016/0167-7799(94)90079-5. [DOI] [PubMed] [Google Scholar]
- Courtney B. C., Williams K. C., Schlager J. J. A phage display vector with improved stability, applicability and ease of manipulation. Gene. 1995 Nov 7;165(1):139–140. doi: 10.1016/0378-1119(95)00526-c. [DOI] [PubMed] [Google Scholar]
- Crameri R., Jaussi R., Menz G., Blaser K. Display of expression products of cDNA libraries on phage surfaces. A versatile screening system for selective isolation of genes by specific gene-product/ligand interaction. Eur J Biochem. 1994 Nov 15;226(1):53–58. doi: 10.1111/j.1432-1033.1994.tb20025.x. [DOI] [PubMed] [Google Scholar]
- Crameri R., Suter M. Display of biologically active proteins on the surface of filamentous phages: a cDNA cloning system for selection of functional gene products linked to the genetic information responsible for their production. Gene. 1993 Dec 27;137(1):69–75. doi: 10.1016/0378-1119(93)90253-y. [DOI] [PubMed] [Google Scholar]
- Gram H., Marconi L. A., Barbas C. F., 3rd, Collet T. A., Lerner R. A., Kang A. S. In vitro selection and affinity maturation of antibodies from a naive combinatorial immunoglobulin library. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3576–3580. doi: 10.1073/pnas.89.8.3576. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jespers L. S., Messens J. H., De Keyser A., Eeckhout D., Van den Brande I., Gansemans Y. G., Lauwereys M. J., Vlasuk G. P., Stanssens P. E. Surface expression and ligand-based selection of cDNAs fused to filamentous phage gene VI. Biotechnology (N Y) 1995 Apr;13(4):378–382. doi: 10.1038/nbt0495-378. [DOI] [PubMed] [Google Scholar]
- Kim K. J., Kanellopoulos-Langevin C., Merwin R. M., Sachs D. H., Asofsky R. Establishment and characterization of BALB/c lymphoma lines with B cell properties. J Immunol. 1979 Feb;122(2):549–554. [PubMed] [Google Scholar]
- Light J., Lerner R. A. Random mutagenesis of staphylococcal nuclease and phage display selection. Bioorg Med Chem. 1995 Jul;3(7):955–967. doi: 10.1016/0968-0896(95)00075-r. [DOI] [PubMed] [Google Scholar]
- Monera O. D., Zhou N. E., Kay C. M., Hodges R. S. Comparison of antiparallel and parallel two-stranded alpha-helical coiled-coils. Design, synthesis, and characterization. J Biol Chem. 1993 Sep 15;268(26):19218–19227. [PubMed] [Google Scholar]
- Rebar E. J., Pabo C. O. Zinc finger phage: affinity selection of fingers with new DNA-binding specificities. Science. 1994 Feb 4;263(5147):671–673. doi: 10.1126/science.8303274. [DOI] [PubMed] [Google Scholar]
- Scott J. K., Smith G. P. Searching for peptide ligands with an epitope library. Science. 1990 Jul 27;249(4967):386–390. doi: 10.1126/science.1696028. [DOI] [PubMed] [Google Scholar]
- Vandamme A. M., Bulens F., Bernar H., Nelles L., Lijnen R. H., Collen D. Construction and characterization of a recombinant murine monoclonal antibody directed against human fibrin fragment-D dimer. Eur J Biochem. 1990 Sep 24;192(3):767–775. doi: 10.1111/j.1432-1033.1990.tb19288.x. [DOI] [PubMed] [Google Scholar]
- Zhou N. E., Kay C. M., Hodges R. S. The role of interhelical ionic interactions in controlling protein folding and stability. De novo designed synthetic two-stranded alpha-helical coiled-coils. J Mol Biol. 1994 Apr 8;237(4):500–512. doi: 10.1006/jmbi.1994.1250. [DOI] [PubMed] [Google Scholar]