POLYCLONAL ANTIBODIES AGAINST MALONDIALDEHYDE-MODIFIED PROTEINS - CHARACTERIZATION AND APPLICATION IN STUDY OF IN-VITRO LIPID-PEROXIDATION OF CELLULAR MEMBRANES

In order to develop an immunofluorescent technique able to detect the cellular damage induced by lipid peroxidation, we produced rabbit poly clonal antibodies (AbAIP) against malondialdehyde (MDA)-modified lysoz yme. These antibodies specifically recognized all the MDA-treated prot eins tested, but not native proteins or proteins treated by other alde hydes. Moreover, we showed by an enzyme-linked immunosorbent assay (EL ISA) that the amount of immunoreactive protein in MDA-treated erythroc ytes was proportional to the concentration of MDA added, suggesting th at this ELISA measurement may represent a quantitative determination o f peroxidative protein alterations. In addition, when coupled to an in direct fluorochrome antibody conjugate [fluorescein isothiocyanate (FI TC)], these antibodies permitted precise localization of the modified proteins within the membranes of peroxidized cells.