重组人肌酸激酶MM同工酶的酶动力学研究

目的:对纯化后重组肌酸激酶MM同工酶(CK-MM)的米氏常数(Km)进行研究,为其作为参考品或校准品应用于临床奠定基础。方法:将重组酶置于自制类人血清基质中,按IFCC参考方法配制试剂,采用HITACHI全自动生化分析仪测定重组酶与天然酶[底物分别是磷酸肌酸(PCr)和腺苷二磷酸(ADP)]的Km常数。结果:重组酶KmPCr值高于天然酶1.4倍,Km ADP重组酶与天然酶近似。结论:重组CK-MM的酶动力学常数基本符合参考方法的测定要求,初步具备作为基因工程人源酶校准品的基本特点。

The kinetic constants study of recombinant human muscle creatine kinase

Objective To study the kinetic constants of the recombinant human muscle creatine kinase(CK-MM) and develop a candidate reference material for CK measurement system. Methods Prepare reagent according to IFCC standard procedure and measure the kinetic constants of the natural creatine kinase in human serum and the recombinant enzyme in self-made matrix with creatine phosphate and ADP as substrate respectively on HITACHI auto-analyzer. Results The kinetic constant of recombinant enzyme is one point four times higher than natural CK with creatine phosphate as substrate and is similar to natural CK with ADP as substrate. Conclusions The kinetic constants of recombinant CK-MM is suitable for determination of reference procedure. The recombinant CK-MM shows primary properties as a calibrator and may be used as a reference material for the determination of serum total creatine kinase.