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Sequence and structure specific antibodies from phage display libraries

J Tanha1, G Forsyth, P Schorr

  • 1Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

Molecular Immunology
|February 1, 1997
PubMed
Summary

Phage display libraries can generate specific antibodies against nucleic acids, but success varies by antigen. Some antibodies recognized single-stranded nucleic acids, while others showed high specificity for double-stranded DNA, demonstrating potential for antibody discovery.

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Area of Science:

  • Molecular Biology
  • Immunology
  • Biochemistry

Background:

  • Phage display technology allows for the selection of antibodies against various targets.
  • Nucleic acid-specific antibodies are valuable tools in molecular biology and diagnostics.
  • The repertoire of antibodies generated by phage display may vary in specificity and representation.

Purpose of the Study:

  • To investigate the feasibility of generating sequence- and structure-specific antibodies against diverse nucleic acid antigens using phage display.
  • To characterize the specificity of antibodies derived from panning against different nucleic acid targets.
  • To assess the representation of antibody sequences within the phage display library.

Main Methods:

  • A large combinatorial phage display library was screened against five nucleic acid antigens: calf thymus DNA, poly[d(GC)], poly[d(AT)], poly(dA) x poly(dT), and poly(rA) x poly(dT).

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  • Antibody clones were selected through multiple rounds of panning.
  • Specificity was evaluated using direct and competitive solid-phase radioimmunoassays.

    • Main Results:

    • Positive clones were obtained against poly[d(GC)], poly(dA) x poly(dT), and poly(rA) x poly(dT) after 3-4 rounds.
    • Clones from poly[d(GC)] panning were non-specific, binding to all nucleic acids.
    • Clones from poly(rA) x poly(dT) showed specificity for single-stranded nucleic acids with sequence preferences.
    • Clones from poly(dA) x poly(dT) exhibited considerable specificity for this antigen.
    • No positive clones were detected for poly[d(AT)] after six rounds, and only two for calf thymus DNA after seven rounds.
    • Sequences of selected antibodies showed no similarity to known DNA-binding antibodies.

    Conclusions:

    • Phage display libraries can yield sequence- and structure-specific antibodies against nucleic acids.
    • The success of antibody recovery is dependent on the specific nucleic acid antigen.
    • The phage display repertoire may not represent all possible antibody sequences, potentially limiting the recovery of antibodies against certain antigens like poly[d(AT)] and calf thymus DNA.