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Sequential analysis by immunoprecipitation-MALDI-TOF: a novel method for detection and identification of alloantibody specificities.

Abstract：

:Alloantibodies are known to influence transplant outcomes. Apart from human leukocyte antigens (HLA), non-HLA targets have been suggested to play a significant role, but little is known about their nature. Here, we present a novel method for identification and characterization of cell surface antigens bound by alloreactive antibodies. Our method consists of 2 consecutive steps: first, immunoprecipitation of cell surface proteins is carried out with serum and, second, matrix-assisted laser desorption/ionization-time-of-flight is used to fingerprint the precipitated cell-surface proteins. As an example, we performed immunoprecipitation with peripheral blood lymphocytes, which had been incubated with an alloreactive serum; immune complexes were coupled to protein-G beads and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; differential protein fractions were then analyzed by matrix-assisted laser desorption/ionization-time-of-flight. The method was validated with serum as well as with plasmapheresis material, which contained antibodies of known HLA specificities, demonstrating its applicability for clinical use.

journal_name

Hum Immunol

journal_title

Human immunology

authors

Heinold A,Kuehl B,Brenner-Weiss G,Opelz G,Tran TH

doi

10.1016/j.humimm.2010.02.006

subject

Has Abstract

pub_date

2010-05-01 00:00:00

pages

462-7

issue

5

eissn

0198-8859

issn

1879-1166

pii

S0198-8859(10)00042-X

journal_volume

71

pub_type

杂志文章

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