Optimal conditions for the separation of rat T lymphocytes on anti-immunoglobulin-immunoglobulin affinity columns

É Kondorosi, J. Nagy, G. Dénes

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The separation of rat T lymphocytes was investigated on anti-IgIg columns. A simple and efficient method for the purification of rat Ig by precipitation of rat serum with sodium sulfate is presented. Protein binding characteristics of glass and plastic beads, as solid support of affinity columns, are described, as well as optimal parameters for coating beads with rat Ig (with BSA, ribonuclease and lysozyme, as comparison). Binding of Ig was primarily dependent on the concentration of the Ig solution. Maximal strong binding of Ig (6.2 × 103 molecules per μ2 of bead surface) was reached at 400 μg per ml concentration of purified Ig solution during 20 min of incubation. Higher concentrations increased only the amount of loosely bound Ig on the surface of beads whereas the amount of firmly bound Ig remained unchanged. Fractionation of lymphoid cell suspensions on anti-IgIg affinity columns prepared at optimal conditions resulted in highly purified T-cell suspensions containing less than 1% of lymphocytes bearing surface Ig receptors.

Original languageEnglish
Pages (from-to)1-13
Number of pages13
JournalJournal of Immunological Methods
Volume16
Issue number1
DOIs
Publication statusPublished - Jun 1977

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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