Preparation of polyclonal antibodies specific for wine proteins

Immunology is an expanding area of research with potentially important appl ications in the analysis of many biological molecules. Polyclonal antibodie s were raised in rabbits against specific proteins as well as against the t otal protein from a Portuguese wine. FPLC cation exchange chromatography wa s used to isolate the total protein fraction and, when in combination with denaturing electrophoresis, to purify individual wine polypeptides. To obta in a high titre, an injection of each antigen followed by three boosters we re given in the immunisation of each rabbit. The titre of the antisera was measured by the ELISA technique and the specificity of the antibodies detec ted by immunoblotting. The antibodies produced were shown to be highly spec ific for the corresponding antigens. However, antibodies obtained specifica lly against a highly purified wine polypeptide seem to recognize the other major wine polypeptides, raising the possibility of structure similarity be tween different wine proteins. Neither the anti-total wine protein antibodi es not the anti-specific wine protein antibodies originated a signal when u sed to probe thaumatin or chitinase. (C) 1999 Society of Chemical Industry.