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Application of thionicotinamide adenine dinucleotide I to homogeneous enzyme immunodiagnostic reagent

A homogeneous enzyme immunization and diagnostic reagent technology, applied in biological testing, material testing products, measuring devices, etc., can solve the problems of poor accuracy and repeatability, poor anti-hemoglobin interference ability, poor stability, etc., to reduce the impact, The effect of reducing detection cost and improving accuracy

Active Publication Date: 2019-06-14
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 thionicotinamide adenine dinucleotide I to homogeneous enzyme immunodiagnostic reagent
  • Application of thionicotinamide adenine dinucleotide I to homogeneous enzyme immunodiagnostic reagent
  • Application of thionicotinamide adenine dinucleotide I to homogeneous enzyme immunodiagnostic reagent

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 an application of a thionicotinamide adenine dinucleotide I to a homogeneous enzyme immunodiagnosticreagent. The diagnostic reagent is a glycocholic acid detection reagent and a CMPF reagent. The defect of relatively poor accuracy and repeatability of a detection result caused by relatively poor stability of the glycocholic acid reagent and the CMPF reagent in the prior art is overcome; a nicotinamide adenine dinucleotide II in the homogeneous enzyme immunodiagnostic reagent for homogeneous enzyme immunoassay is replaced with the thionicotinamide adenine dinucleotide I, so that the problem of relatively poor stability of the homogeneous enzyme immunodiagnostic reagent is effectively solved,the repeatability of the detection result of the reagent is improved, and the accuracy of the detection result is improved; and meanwhile, the anti-hemoglobin interference resistance of the reagent isimproved, the influence of a hemolysis phenomenon of a clinical sample on the detection result is weakened, and the detectability of the homogeneous enzyme immunodiagnostic reagent is improved, so that the test reagent is suitable for a conventional microplate reader, the detection cost is effectively reduced, and meanwhile, the usage amount of the reagent and raw materials is reduced.

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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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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