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Preparation and mass spectrometric study of egg yolk antibody (IgY) against rabies virus
Author(s) -
Sun Shuqing,
Mo Wenjun,
Ji Yiping,
Liu Shuying
Publication year - 2001
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.271
Subject(s) - chemistry , chromatography , molecular mass , rabies virus , virus , size exclusion chromatography , dithiothreitol , yolk , antigen , antibody , virology , biochemistry , enzyme , food science , biology , genetics , immunology
Rabies virus was used as the antigen to immunize laying chickens. Anti‐rabies virus immunoglobulin Y(IgY) was isolated from yolks of the eggs laid by these chickens using a two‐step salt precipitation and one‐step gel filtration protocol. The purified IgY was reduced with dithiothreitol, and heavy chains (HC) and light chains (LC) were obtained. In addition, the purified IgY was digested with pepsin and the fragment with specific antigen binding properties (Fab) was produced. Using matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐TOFMS), the average molecular weights of IgY, HC, LC, and Fab were determined as 167 250, 65 105, 18 660, and 45,359 Da, respectively. IgY has two structural differences compared with mammalian IgGs. First, the molecular weight of the heavy chain of IgY is larger than that of its mammalian counterpart, while the molecular weight of the light chain of IgY is smaller. Second, upon pepsin digestion, anti‐rabies virus IgY is degraded into Fab, in contrast to mammalian IgG, which has been reported to be degraded into F(ab′) 2 under the same conditions. Copyright © 2001 John Wiley & Sons, Ltd.