Assessment of a positive selection technique using an avidin column to isolate human peripheral blood T cell subsets

Laurie S. Davis, Melissa J. McIlraith, Trang Paecheo, Brent Becker, Leah M. Adix, Ranjeny Thomas, Mary C. Wacholtz, Peter E. Lipsky

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

The current studies were designed to assess a new technique for positively selecting human T cells from whole peripheral blood mononuclear cells using the minimal amount of monoclonal antibody required to bind the T cell to an avidin column indirectly via a biotin-conjugated secondary antibody. Positive selection of T cells has previously been avoided because the saturating amounts of antibodies required for other isolation procedures can lead to aberrant results in assay of T cell activation and function. The avidin column technique for obtaining purified T cell subsets was compared to a multi-step procedure that included negative selection panning. The positive selection technique was easily performed within 4 h whereas the negative selection technique required a minimum of 12 h to complete. The avidin column technique proved to be a rapid and simple method for isolating T cell subsets of high purity and normal functional capabilities. Since minimal amounts of monoclonal antibodies were used for the purification protocol, no consistent inhibitory or stimulatory effect of the residual antibody was noted in assays of activation and proliferation of positively selected T cells compared to T cells isolated by negative selection panning.

Original languageEnglish (US)
Pages (from-to)247-257
Number of pages11
JournalJournal of Immunological Methods
Volume175
Issue number2
DOIs
StatePublished - Oct 14 1994

Keywords

  • (Human)
  • Activation and function
  • Avidin column
  • Positive and negative selection
  • T lymphocyte

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Davis, L. S., McIlraith, M. J., Paecheo, T., Becker, B., Adix, L. M., Thomas, R., Wacholtz, M. C., & Lipsky, P. E. (1994). Assessment of a positive selection technique using an avidin column to isolate human peripheral blood T cell subsets. Journal of Immunological Methods, 175(2), 247-257. https://doi.org/10.1016/0022-1759(94)90367-0