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Detection of immune complexes by matrix‐assisted laser desorption/ionization mass spectrometry
Author(s) -
Schlosser Gitta,
Pocsfalvi Gabriella,
Malorni Antonio,
Puerta Angel,
de Frutos Mercedes,
Vékey Károly
Publication year - 2003
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1239
Subject(s) - chemistry , mass spectrometry , desorption , matrix (chemical analysis) , mass spectrometry imaging , ionization , analytical chemistry (journal) , chromatography , direct electron ionization liquid chromatography–mass spectrometry interface , chemical ionization , ion , organic chemistry , adsorption
Matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS) was used to detect an immune complex formed between β ‐lactoglobulin and polyclonal anti‐ β ‐lactoglobulin antibody in the gas phase. The most important experimental parameters to detect such a specific antibody–antigen complex by MALDI were the use of solutions at near‐neutral pH and of sinapinic acid matrix prepared by the dried‐droplet method. Under such conditions, predominantly one but also two molecules of antigen protein were complexed by the antibody. Specific formation of the antibody–antigen complex was confirmed by performing competitive reactions. Addition of antibody to a 1:1 mixture of β ‐lactoglobulin and one control protein resulted not only in the appearance of the expected antibody–antigen complex, but also in a strong decrease in the free β ‐lactoglobulin signal, while the abundance of the control protein was not influenced. Copyright © 2003 John Wiley & Sons, Ltd.