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Preparation of polyclonal antibodies specific for wine proteins
Author(s) -
Monteiro Sara,
PiçarraPereira Maria A,
Tanganho Maria Conceição,
Rente José Pedro,
Loureiro Virgílio B,
Teixeira Artur R,
Ferreira Ricardo B
Publication year - 1999
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/(sici)1097-0010(199904)79:5<772::aid-jsfa251>3.0.co;2-j
Subject(s) - thaumatin , polyclonal antibodies , wine , fast protein liquid chromatography , antibody , antiserum , antigen , oenology , chemistry , titer , biochemistry , affinity chromatography , microbiology and biotechnology , biology , immunology , food science , enzyme , viticulture , gene
Immunology is an expanding area of research with potentiallyimportant applications in the analysis of many biological molecules.Polyclonal antibodies were raised in rabbits against specificproteins as well as against the total protein from a Portuguese wine.FPLC cation exchange chromatography was used to isolate the totalprotein fraction and, when in combination with denaturingelectrophoresis, to purify individual wine polypeptides. To obtain ahigh titre, an injection of each antigen followed by three boosterswere given in the immunisation of each rabbit. The titre of theantisera was measured by the ELISA technique and the specificity ofthe antibodies detected by immunoblotting. The antibodies producedwere shown to be highly specific for the corresponding antigens.However, antibodies obtained specifically against a highly purifiedwine polypeptide seem to recognize the other major wine polypeptides,raising the possibility of structure similarity between differentwine proteins. Neither the anti‐total wine protein antibodiesnot the anti‐specific wine protein antibodies originated asignal when used to probe thaumatin or chitinase. © 1999 Society of Chemical Industry