Application of oxidized thio-coenzyme I in homogeneous enzyme immunodiagnostic reagents

A homogeneous enzyme immunoassay and diagnostic reagent technology, applied in biological testing, instruments, measuring devices, etc., can solve the problems of poor anti-hemoglobin interference, poor accuracy and repeatability, poor stability, etc., to reduce detection costs and weaken Influence, the effect of improving accuracy

Active Publication Date: 2022-03-08
HANGZHOU BOPU MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the poor stability of the oxidized coenzyme II (NADP) contained in the glycochlic acid reagent and the CMPF reagent in the prior art, which leads to the poor accuracy and repeatability of the overall reagent in the detection results. And the defect of poor ability to resist hemoglobin interference
[0009] Therefore, the first object of the present invention is to effectively solve the problem of homogeneous enzyme immunoassay by replacing the oxidized coenzyme II (NADP) in the homogeneous enzyme immunodiagnostic reagent with oxidized sulfo-coenzyme I (coenzyme analogue). Diagnose the problem of poor stability of reagents, improve the repeatability of reagent test results, and improve the accuracy of test results

Method used

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  • Application of oxidized thio-coenzyme I in homogeneous enzyme immunodiagnostic reagents
  • Application of oxidized thio-coenzyme I in homogeneous enzyme immunodiagnostic reagents
  • Application of oxidized thio-coenzyme I in homogeneous enzyme immunodiagnostic reagents

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Preparation of Glycocholic Acid Reagent:

[0047] (1) Preparation of reagent R1 and reagent R2

[0048] (1-1) Accurately weigh 1 L of prepared (30-70) mM tris buffer, fully dissolve and mix well, and adjust the pH to 6.0-7.0;

[0049] (1-2) Take 800 ml of buffer solution in step 1-1, add (4-5) mM G6P, 0.05% pc-300, (1.2-2.0) KU / L glycocholic acid monoclonal antibody successively, fully Dissolve and mix well for later use;

[0050] (1-3) Take 400ml of the solution in step 1-2, add (4-6)mM oxidized thio-coenzyme I, mix evenly, keep it at 4°C for overnight use, and mark it as R1;

[0051] (1-4) Take 200ml of buffer solution in step 1-1, add (0.15-0.2)% BSA, (1.5-2)% NaCl, 0.05% PC-300, (0.5-0.6) KU / L glucose hexaphosphate in sequence The dehydrogenase-glycocholic acid conjugate is labeled R2.

[0052] (2) Preparation of calibrators and quality control products

[0053] (2-1) Weigh (0.7-0.9)% NaCl, (0.05-0.06)% pc-300, dissolve in purified water, and prepare as a speci...

Embodiment 2

[0092] Preparation of CMPF reagent:

[0093] (1) Preparation of reagent R1 and reagent R2

[0094] (1-1) Accurately weigh 1 L of prepared (30-70) mM tris buffer, fully dissolve and mix well, and adjust the pH to 6.0-7.0;

[0095] (1-2) Take 800ml of the buffer in step 1-1, add (4-5)mM G6P, 0.05% pc-300, (1.2-2.0)KU / L CMPF antibody in sequence, fully dissolve and mix well for backup use;

[0096] (1-3) Take 400ml of the solution in step 1-2, add (4-6)mM oxidized thio-coenzyme I, mix evenly, keep it at 4°C for overnight use, and mark it as R1;

[0097] (1-4) Take 200ml of buffer solution in step 1-1, add (0.15-0.2)% BSA, (1.5-2)% NaCl, 0.05% PC-300, (0.5-0.6) KU / L glucose hexaphosphate in sequence The dehydrogenase-CMPF conjugate is labeled R2.

[0098](2) Preparation of calibrators and quality control products

[0099] (4-1) Weigh (0.7-0.9)% NaCl, (0.05-0.06)% pc-300, dissolve with purified water, and prepare it as a special diluent for CMPF calibration quality control pro...

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Abstract

The invention relates to the technical field of in vitro diagnosis and detection, in particular to the application of oxidized thio-coenzyme I in homogeneous enzyme immunodiagnostic reagents, and the diagnostic reagents are glycocholic acid detection reagents and CMPF reagents. The present invention overcomes the poor stability of glycocholic acid reagent and CMPF reagent in the prior art, resulting in the defects of poor detection result accuracy and repeatability, by combining the oxidative Type coenzyme II is replaced by oxidized thio-coenzyme I, which effectively solves the problem of poor stability of homogeneous enzyme immunodiagnostic reagents, improves the repeatability of reagent test results, and improves the accuracy of test results; at the same time, it improves the anti-hemoglobin of the reagent. Interference ability, weaken the influence of clinical sample hemolysis on the test results, improve the detectability of homogeneous enzyme immunodiagnostic reagents, make the test reagents suitable for conventional microplate readers, effectively reduce the cost of testing, and save the amount of reagents and raw materials.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis and detection, in particular to the application of oxidized thio-coenzyme I in homogeneous enzyme immunodiagnostic reagents. Background technique [0002] NADP is the abbreviation of nicotinamide adenine dinucleotide phosphate (nicotinamide adenine dinucleotidephosphate), which was once called pyridine nucleotide triphosphate (TPN) or co-dehydrogenase II or oxidized coenzyme II. It is a substance in which nicotinamide adenine dinucleotide and a phosphate molecule are bonded by an ester bond, and is widely found in the biological world. Its chemical properties, absorption spectrum and redox form are similar to nicotinamide adenine dinucleotide (NAD). [0003] Nicotinamide adenine dinucleotide (NAD)-dependent oxidoreductase has important physiological functions, but it is difficult to selectively study; at the same time, the corresponding redox reaction needs to consume stoichiometric co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/577
Inventor 王轶雄程小龙余琳邓光兴
Owner HANGZHOU BOPU MEDICAL TECH
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