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Related Experiment Videos

Screening cDNA expression libraries in lambda gt11 with a T cell hybridoma.

S Gillard-Blaas1, M Ayane, C Wirbelauer

  • 1Max-Planck-Institut für Immunobiologie, Freiburg, F.R.G.

Journal of Immunological Methods
|June 3, 1991
PubMed
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This study presents a novel method for direct T cell screening of Plasmodium chabaudi chabaudi cDNA libraries. The technique efficiently identifies T cell epitopes, aiding in the discovery of crucial peptides for infectious diseases.

Area of Science:

  • Immunology
  • Molecular Biology
  • Parasitology

Background:

  • T cell hybridomas are crucial for studying T cell responses.
  • Screening cDNA libraries is essential for identifying pathogen antigens.
  • Plasmodium chabaudi chabaudi causes malaria, necessitating new diagnostic and therapeutic targets.

Purpose of the Study:

  • To develop and validate a direct T cell screening system for Plasmodium chabaudi chabaudi cDNA libraries.
  • To identify T cell epitopes recognized by specific T cell hybridomas.
  • To assess the sensitivity and applicability of the screening technique.

Main Methods:

  • Utilized a T cell hybridoma specific for P. chabaudi chabaudi proteins.
  • Employed a lambda gt11 expression cDNA library of P. chabaudi chabaudi.

Related Experiment Videos

  • Developed a method using antibody-coated polystyrene beads for recombinant fusion protein separation.
  • Co-cultured beads with antigen-presenting cells and T cell hybridoma for screening.

    • Main Results:

    • The system successfully screened the P. chabaudi chabaudi cDNA library.
    • The technique demonstrated high sensitivity, detecting one positive phage in 1000-10,000.
    • Enabled selection of individual positive phage plaques through serial dilutions.
    • The method proved effective in identifying T cell-reactive clones.

    Conclusions:

    • A sensitive and efficient direct T cell screening system for cDNA libraries was established.
    • This technique is broadly applicable for identifying T cell epitopes in infectious diseases.
    • The method facilitates the discovery of key T cell peptides for vaccine and therapeutic development.