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Cross-linked filamentous phage as an affinity matrix

G P Smith1, V A Petrenko, L J Matthews

  • 1Division of Biological Sciences, University of Missouri, Columbia 65211, USA. gpsmith@biosci.mbp.missouri.edu

Journal of Immunological Methods
|September 23, 1998
PubMed
Summary

Filamentous phage can be cross-linked into stable aggregates for antibody purification. This method streamlines research by creating an affinity matrix directly from selected phage displaying peptide epitopes.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Virology

Background:

  • Filamentous phage are versatile biological tools.
  • Immunoaffinity purification often requires robust matrices.
  • Phage display libraries are used for peptide epitope discovery.

Purpose of the Study:

  • To develop a streamlined method for antibody purification using filamentous phage.
  • To create a stable affinity matrix from filamentous phage displaying peptide epitopes.

Main Methods:

  • Cross-linking filamentous phage to form a hydrophilic aggregate.
  • Pelleting the aggregate via low-speed centrifugation.
  • Genetically fusing a peptide epitope to a coat protein on the phage surface.
  • Utilizing the phage aggregate as an affinity matrix for antibody absorption.

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Main Results:

  • The filamentous phage aggregate is stable at near-neutral pH and acid buffers (pH 2.2).
  • The aggregate effectively absorbs and purifies antibodies that bind to the displayed peptide epitope.
  • This method simplifies the process of creating affinity matrices from selected phage.

Conclusions:

  • Cross-linked filamentous phage aggregates provide a stable and effective platform for antibody purification.
  • This technique significantly streamlines research and development in antibody discovery and purification.
  • Directly using selected phage to create affinity matrices offers a considerable advantage in laboratory workflows.