PMID: 2121829Oct 19, 1990Paper

Direct blotting with viable cells of protein mixtures separated by two-dimensional gel electrophoresis

Journal of Immunological Methods
H GulleS H Kaufmann

Abstract

A procedure is described which combines the high resolution power of two-dimensional (2D) gel electrophoresis with the advantage of direct probing with viable cells. This device permits the transfer by electroelution of 480 distinct fractions from a 2D gel into soluble phase. Transferred fractions are virtually nontoxic, thus allowing direct probing with viable cells. Using this procedure it was shown that T cells from normal healthy individuals recognized a multitude of Mycobacterium tuberculosis antigens and that the fine antigen recognition pattern of T cells changed after short-term culture in vitro. The application of this procedure to the verification of antigen purity at the T cell level and to the identification of antigens within crude bacterial lysates which are recognized by cloned T cells is described. This approach should be applicable to the rapid identification and characterization of any interesting T cell antigen, for example from important pathogens against which a subunit vaccine is desirable. Moreover, it could be helpful for the analysis of interactions between soluble ligands and their target cells.

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Citations

Aug 1, 1996·Molecular Aspects of Medicine·M J Colston
Sep 1, 1995·Veterinary Immunology and Immunopathology·H GulleK M Moriarty
Jan 5, 2002·Molecular and Biochemical Parasitology·Gaby ReichmannHans-Georg Fischer
Apr 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·T H KongT M Shinnick
Jan 1, 1992·Scandinavian Journal of Immunology. Supplement·S H KaufmannB Schoel
Aug 1, 1990·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·M E MunkS H Kaufmann
Jul 2, 1998·Clinical and Experimental Immunology·G FörsterG Kahaly
Nov 16, 2011·ISRN Surgery·Davide BlonnaFilippo Castoldi
Feb 14, 1992·Analytical Biochemistry·A Persidis, A A Harcombe
Oct 25, 2003·Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Médicas E Biológicas·A AlitoA Cataldi
Apr 29, 2000·Electrophoresis·P Cash
Mar 1, 1997·FEMS Immunology and Medical Microbiology·B A DegnanJ A Goodacre

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