COMPARISON OF TWO METHODS FOR CANINE C-REACTIVE PROTEIN (CRP) ISOLATION AND PURIFICACTION
Abstract
C-reactive protein (CRP) is one of the major acute phase proteins in dogs with a pronounced and rapid increase in response to infection and tissue injury. Canine CRP analysis has demonstrated to be useful in many clinical and experimental situations for evaluating, diagnosing and monitoring several inflammatory processes. Purified CRP could be used as a calibrator in immunoassays or as an antigen for producing anti-canine CRP antibodies. In the present study CRP was isolated from canine acute phase serum by using two different techniques: 1. Immunopurification, by using polyclonal anti-CRP antibodies bound to CNBr-sepharose. 2. Affinity chromatography on agarose coupled with phosphorylethanolamine through epoxy bound. The purity of the purified proteins with both methodologies was assessed by sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE). Both methods could be applied for canine CRP purification, however affinity chromatography provided a more purified protein and in bigger amount than immunopurification did and besides constitutes a cheaper option since antibodies, which represent a very expensive stuff, are not needed.Downloads
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