Preparation of polyclonal antibodies specific for wine proteins

✍ Scribed by Monteiro, Sara; Piçarra-Pereira, Maria A; Tanganho, Maria Conceição; Rente, José Pedro; Loureiro, Virgílio B; Teixeira, Artur R; Ferreira, Ricardo B

• Publisher: John Wiley and Sons
• Year: 1999
• Tongue: English
• Weight: 246 KB
• Volume: 79
• Category: Article
• ISSN: 0022-5142
• DOI: 10.1002/(sici)1097-0010(199904)79:5<772::aid-jsfa251>3.0.co;2-j

✦ Synopsis

Immunology is an expanding area of research with potentially important applications in the analysis of many biological molecules. Polyclonal antibodies were raised in rabbits against speciüc proteins as well as against the total protein from a Portuguese wine. FPLC cation exchange chromatography was used to isolate the total protein fraction and, when in combination with denaturing electrophoresis, to purify individual wine polypeptides. To obtain a high titre, an injection of each antigen followed by three boosters were given in the immunisation of each rabbit. The titre of the antisera was measured by the ELISA technique and the speciücity of the antibodies detected by immunoblotting. The antibodies produced were shown to be highly speciüc for the corresponding antigens. However, antibodies obtained speciücally against a highly puriüed wine polypeptide seem to recognize the other major wine polypeptides, raising the possibility of structure similarity between diþ erent wine proteins. Neither the anti-total wine protein antibodies not the anti-speciüc wine protein antibodies originated a signal when used to probe thaumatin or chitinase.