MONOCLONAL ANTIBODIES FOR AN ELISA FOR THE ETHYLENE-RELATED PROTEIN PAPER-1

NEOGEN CORPORATION HAS DEVELOPED NUMEROUS IMMUNOASSAYS CURRENTLY IN WIDE COMMERCIAL PRACTICE TO DETECT AND QUANTIFY TOXIC RESIDUES (E.G. MYCOTOXINS, SULFONAMIDES). TECHNOLOGY IS CURRENTLY BEING DEVELOPED FOR DETECTING AND QUANTIFYING THE DEVELOPMENTALLY REGULATED ENZYME 1-AMINOCYCLOPROPANE-1-CARBOXYLATE (ACC) OXIDASE IN ETHYLENE BIOSYNTHESIS IN PLANTS, EXPRESSED IN CLIMACTERIC FRUITS AS THEY MATURE AND OBLIGATORY FOR RIPENING. AN ACC OXIDASE POLYCLONAL ANTIBODY-BASED ELISA WAS DEVELOPED BY NEOGEN CORPORATION FOR RESEARCH AS A MATURITY/RIPENING INDEX FOR CLIMACTERIC FRUITS. WE HAVE FOUND THAT AN ETHYLENE-RELATED PROTEIN. PAPER-1, IS EXPRESSED IN FRUITS IN RESPONSE TO EXOGENOUS ETHYLENE JUST BEFORE THE EXPRESSION OF ACC OXIDASE. THIS PROTEIN MAY REGULATE THE EXPRESSION OF ACC OXIDASE VIA A POSITIVE FEED-BACK CONTROL MECHANISM, AS SUCH, THE DETECTION OF PAPER-1 IN FRUITS WOULD PRE-STAGE THE APPEARANCE OF ACC OSIDASE AND PROVIDE A MEANS TO ALERT FRUIT GROWERS THAT FRUIT MATURATION HAS BEEN COMPLETED AND THAT THE ONSET OF RIPENING IS EMINENT. THE SPECIFIC OBJECTIVE OF THIS PROPOSAL IS TO PRODUCE AND CHARACTERIZE MONOCLONAL ANTIBODIES (MAB) AGAINST THE PAPER-1 ANTIGEN. THESE MABS WILL SUBSEQUENTLY BE USED TO DEVELOP AN ELISA TO DETECT AND QUANTIFY PAPER-1 AS AN INDEX FOR USE ALONE OR IN TANDEM WITH THE ACC OXIDASE ELISA, THE LATTER ASSESSING STATUS OF RIPENING.