Detection of immune complexes by matrix-assisted laser desorption/ionization mass spectrometry |
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Authors: | Schlosser Gitta Pocsfalvi Gabriella Malorni Antonio Puerta Angel de Frutos Mercedes Vékey Károly |
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Affiliation: | Chemical Research Center, Hungarian Academy of Sciences, Budapest, Hungary. |
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Abstract: | Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was used to detect an immune complex formed between beta-lactoglobulin and polyclonal anti-beta-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 beta-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 beta-lactoglobulin signal, while the abundance of the control protein was not influenced. |
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