CPC C12Q 1/32 (2013.01) [C12Q 1/008 (2013.01); C12Q 1/68 (2013.01); G01N 21/78 (2013.01); G01N 33/53 (2013.01); G01N 33/535 (2013.01); G01N 33/5695 (2013.01); G01N 33/581 (2013.01); G01N 21/25 (2013.01); G01N 2333/35 (2013.01); G01N 2333/90209 (2013.01)] | 3 Claims |
1. An enzymatic measurement method using an antibody-enzyme complex, the method being implemented in which an enzyme reaction product of the antibody-enzyme complex is assayed by generating thio-NADH and/or thio-NADPH by an enzymatic cycling reaction using NADH and/or NADPH, thio-NAD and/or thio-NADP and a dehydrogenase (DH), followed by:
i) measuring the generated thio-NADH and/or thio-NADPH, or
ii) measuring a color change caused by the generated thio-NADH and/or thio-NADPH, wherein
an enzyme reaction system that selectively reduces the NAD and/or NADP generated from NADH and/or NADPH by the enzymatic cycling reaction is included in the enzymatic cycling reaction system,
wherein the enzyme reaction system that selectively reduces the NAD and/or NADP uses a substrate that does not become a substrate for the enzyme of the antibody-enzyme complex and a substrate for the enzyme of the enzymatic cycling reaction, and an enzyme that does not react with the substrate for the enzyme of the antibody-enzyme complex and the substrate of the enzyme of the enzymatic cycling reaction; and
wherein the combination of enzyme and substrate used in the enzyme reaction system that selectively reduces NAD and/or NADP is selected from the group consisting of:
(i) glutamate dehydrogenase and glutamic acid;
(ii) leucine dehydrogenase and leucine;
(iii) alanine dehydrogenase and alanine;
(iv) phenylalanine dehydrogenase and phenylalanine;
(v) serine dehydrogenase and serine;
(vi) valine dehydrogenase and valine;
(vii) tryptophan dehydrogenase and tryptophan; and
(viii) aspartate dehydrogenase and aspartic acid.
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