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Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci pp 241–249Cite as

Cell-based panning as a means to isolate phage display Fabs specific for a bacterial surface protein

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Abstract

Surface proteins provide a multitude of functions for the bacterial cell. Antibodies to these proteins can provide tools for tagging bacteria and characterizing protein function. Phage display technology has emerged as a powerful method for producing monoclonal Fabs in Escherichia coli. In an effort to study the adhesion mechanisms of Streptococcus parasanguis FW213, Fabs specific for the surface adhesin protein Fap1 were produced using phage display. The immune repertoire of a mouse injected with purified Fapl was cloned into the phagemid vector pCOMB3, and a combinatorial Fab library was expressed in E. coli. A cell-based panning method using whole S. parasanguis cells was developed and has been shown to be a means for enriching for Fabs specific for the Fapl protein.

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Abbreviations

Fab:

fragment antigen binding

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Author information

Authors and Affiliations

  1. Department of Microbiology and Molecular Genetics, University of Vermont, Burlington, Vermont, USA

    Aimee E. Stephenson & Robert J. Melamede

  2. Department of Microbiology and Molecular Genetics, College of Medicine and College of Agriculture and Life Sciences, Stafford Hall, University of Vermont, Burlington, Vermont, 05405, USA

    Paula Fives-Taylor

Authors
  1. Aimee E. Stephenson
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  2. Paula Fives-Taylor
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  3. Robert J. Melamede
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Corresponding author

Correspondence to Paula Fives-Taylor .

Editor information

Editors and Affiliations

  1. Department of Microbiology and Molecular Genetics, University of Vermont, Stafford Building, 05405, Burlington, Vermont, USA

    Paula M. Fives-Taylor

  2. Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana, USA

    Donald J. LeBlanc

  3. Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, IN, USA

    Donald J. LeBlanc

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© 1998 Springer Science+Business Media Dordrecht

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Stephenson, A.E., Fives-Taylor, P., Melamede, R.J. (1998). Cell-based panning as a means to isolate phage display Fabs specific for a bacterial surface protein. In: Fives-Taylor, P.M., LeBlanc, D.J. (eds) Methods for studying the genetics, molecular biology, physiology, and pathogenesis of the streptococci. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-2258-2_27

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  • DOI: https://doi.org/10.1007/978-94-017-2258-2_27

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-90-481-5262-9

  • Online ISBN: 978-94-017-2258-2

  • eBook Packages: Springer Book Archive

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