| Abstract: | A panel of four monoclonal antibodies (Mabs) was raised against β-conglycinin, the 7S globulin from soya bean. The antibodies were characterised by using direct and competitive enzyme-linked immunosorbent assays (ELISAs), immunoblotting procedures and by analysis of subunit fractions obtained after anion exchange chromatography. All the Mabs were specific for β-conglycinin, recognising the acidic α-and α-subunits. One of the Mabs (IRFN 0089) was used to probe the structural changes that take place during thermal denaturation of β-conglycinin. A two-site ELISA was developed to observe structural changes in β-conglycinin with heating. Antibody recognition of heated conglycinin increased with temperature reaching a maximum at 65°C; β-conglycinin heated to this temperature was recognised three-fold better than unheated β-conglycinin. At a higher temperature (65–95°C) β-conglycinin immunoreactivity remained at least two-fold higher than that of the unheated protein. Differential scanning calorim-etry data showed the maximum binding of Mab 0089 to correspond with the thermal transition of the β-conglycinin molecule. The use of the panel of antibodies as structural probes is discussed together with further possible applications of this technology in food chemistry. |
