| 摘要 |
Disclosed is a kit for performing a sequential "Specific Immunological Extraction Followed by Enzymatic Detection" (SIEFED)- "Enzyme-Linked Immuno Sorbent Assay" (ELISA) measuring an activation status of an enzyme or cells which may release the said enzyme in a biological sample obtained from a mammal or measuring effects of synthethized or natural compounds on the said enzyme, and which comprises: - one solid support surface, - a primary antibody or hypervariable portion thereof, both specific of the said enzyme and being fixed upon said solid support surface, - a suitable substrate to be transformed by the enzyme into a visible reaction product, - a secondary antibody or a hypervariable portion thereof both being specific of the said enzyme, wherein the first antibody and the second antibody do not cross react between them, - reagents for a detection of a complex of same said primary antibody -said enzyme -said secondary antibody fixed on the same said solid support surface, and - a sample of the said pure enzyme with a determined enzymatic activity and a determined total quantity, to be used for establishing SIEFED and ELISA reference curves. Also disclosed is an in vitro method for a measurement of an enzymatically active fraction and a total amount of an enzyme in a same complex biological sample, said method being a SIEFED method followed by an ELISA method, this "combined SIEFED-ELISA" method being performed sequentially with the same primary antibody or hypervariable portion thereof both being specific of the said enzyme, which is fixed upon the same solid support and wherein the SIEFED method and ELISA method are performed sequentially upon the same solid support using the same primary antibody.
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