Monoclonal antibodies as probes for differentiating tropoelastin a and b

BALB/C mice were immunized with a mixture of aortic tropoelastin a and b. To ensure the presence of tropoelastin a, the lathyritic chick aortic tissue was first extracted with denaturing and reducing agents. After dialysis, the standard procedure for tropoelastin purification was followed. Following immunization, the lymphocytes were fused with a myeloma cell line. Resulting hybridomas were cloned and tested for reactivity towards tropoelastin a and b. The majority of monoclonal antibodies exhibited crossreactivity between the two tropoelastins. Two of the clones were further characterized because of their ability to differentiate both tropoelastin a and b. As demonstrated by immunoprecipitation and Western blot analyses, one clone reacted preferentially with tropoelastin b and the other with tropoelastin a and b. Significantly, these results demonstrate that tropoelastin a and b can be differentiated by immunological techniques.