Preparation of polyclonal antibodies specific for wine proteins

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 specific 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 specificity of the antibodies detected by immunoblotting. The antibodies produced were shown to be highly specific for the corresponding antigens. However, antibodies obtained specifically against a highly purified wine polypeptide seem to recognize the other major wine polypeptides, raising the possibility of structure similarity between different wine proteins. Neither the anti-total wine protein antibodies not the anti-specific wine protein antibodies originated a signal when used to probe thaumatin or chitinase.