Rapid separation of plasma creatine kinase isoenzymes

Abstract

An improved method for the rapid separation of the MM and MB creatine kinase isoenzymes is provided. The method involves the use of an ion exchange support material comprising porous glass beads having a stationary phase coupled to the surfaces thereof through an intermediate coupling agent. The intermediate coupling agent is preferably a silane coupling agent constituted by an organosilane with a silicon functional group capable of bonding to the surfaces of the glass beads and an organic functional group capable of bonding to the stationary phase. Less preferably, the coupling agent may be formed from a Grignard reagent of the formula RMgX where R is lower alkenyl and X is halogen. A blood plasma or serum sample and a buffer of low ionic strength are contacted with the ion exchange support, the resultant mixture is incubated to effect MB isoenzyme adsorption by the support, the supernatant liquid fraction containing the MM isoenzyme is separated from the support, the support is washed to effect a removal of residual of MM therefrom, a buffered solution of a strong electrolyte is added to the support to effect desorption of the MB isoenzyme therefrom and the supernatant liquid fraction containing the MB isoenzyme is separated from the support. The MM and MB activity in the respective liquid fractions thus obtained is then assayed to provide an index of myocardial damage.

Dependencies are inferred from the claim text.