Sequential analysis by immunoprecipitation-MALDI-TOF: a novel method for detection and identification of alloantibody specificities
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 antige...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
21 February 2010
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| In: |
Human immunology
Year: 2010, Volume: 71, Issue: 5, Pages: 462-467 |
| ISSN: | 1879-1166 |
| DOI: | 10.1016/j.humimm.2010.02.006 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/j.humimm.2010.02.006 Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/abs/pii/S019888591000042X?via%3Dihub 
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| Author Notes: | Andreas Heinold, Boris Kuehl, Gerald Brenner-Weiss, Gerhard Opelz, Thuong Hien Tran |
| Summary: | 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. |
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| Item Description: | Gesehen am 02.06.2021 |
| Physical Description: | Online Resource |
| ISSN: | 1879-1166 |
| DOI: | 10.1016/j.humimm.2010.02.006 |