Antiserum was prepared by immunizing SPF-grade Balb/C mice with the whole-cell protein of the highly virulent Listeria monocytogenes 819-2 strain.The extracellular proteome of the strain was analyzed using the immunoproteomics approach,to identify and obtain strong immunogenic proteins of L.monocytogenes as candidate antigens for specific antibody preparation.The whole-cell protein of the 819-2 strain was extracted using the ultrasonic method.The SPF-grade Balb/C mice were immunized four times using this protein extract to prepare antiserum,and the titer of the antiserum equaled 1:512 000 as measured using indirect ELISA.The extracellular protein of L.monocytogenes 819-2 strain was extracted using TCA precipitation with 10%(m/V)deoxycholate sodium(DOC).The spots for strong immunogenic proteins were identified with the help of immunoproteomics and LC-MS/MS.Two-dimensional electrophoresis results showed 85 protein spots and five strong immunogenic proteins,including P60,InlC,MltG,Enolase,and YxeA family protein,were successfully identified.The results provide a foundation for the preparation of specific antibodies based on highly immunogenic proteins for the enrichment and rapid detection of L.monocytogenes in food and food processing environments.
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