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Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments

Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments

DOI:
10.3791/54583-v

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12:28 min

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October 15, 2016

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, , , ,
1Fischell Department of Bioengineering,University of Maryland, 2Department of Chemical and Biomolecular Engineering,University of Maryland

Chapters

  • 00:05Title
  • 01:11Express scFv Library and Prepare Spheroplasts
  • 03:46Immobilize Target Antigen onto Magnetic Beads
  • 05:05Screen scFv Library
  • 06:32Identify Clones in Secondary Screen Using ELISA
  • 09:51Results: scFv Antibody Library Screening Using the E. coli Twin-arginine Translocation Pathway
  • 11:39Conclusion

Summary

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Automatic Translation

We provide a method to simultaneously screen a library of antibody fragments for binding affinity and cytoplasmic solubility by using the Escherichia coli twin-arginine translocation pathway, which has an inherent quality control mechanism for intracellular protein folding, to display the antibody fragments on the inner membrane.

Tags

Keywords: Antibody FragmentInner-membrane DisplayTwin-arginine TranslocationAntibody EngineeringCytoplasmic FoldingSpheroplastsScFv LibraryFractionation BufferLysozymeEDTAMgCl2

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