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Composition and application of composition as enzyme labeled compound preserving fluid

A composition and selected technology, which are applied in the directions of chemiluminescence/bioluminescence, analysis by chemical reaction of materials, and measurement devices, which can solve the problems of unsatisfactory stabilization effect of enzyme-labeled complexes and inability to meet stability requirements. , to achieve the effect of improving stability and prolonging shelf life

Inactive Publication Date: 2017-05-31
SINOCARE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in order to maintain the stability of the enzyme-labeled complex, it is usually necessary to use a buffer with a stabilizing function. However, the existing buffer has an unsatisfactory effect on the stabilization of the enzyme-labeled complex, and usually can only maintain the enzyme-labeled complex in a short period of time. The activity of the complex is far from meeting the stability requirements

Method used

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  • Composition and application of composition as enzyme labeled compound preserving fluid
  • Composition and application of composition as enzyme labeled compound preserving fluid
  • Composition and application of composition as enzyme labeled compound preserving fluid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] 1. Configure storage solution

[0092] 1L specific formula: MES 3.91g, NaCl 8.8g, KCl 0.2g, aminopyrine 1g, BSA 2.5g, casein 2.5g, sucrose 20g, glycerol 100mL, Tween 20, 0.5mL, Proclin300, 0.5mL. Use sterilized water to prepare, adjust the pH to 6.2, filter with 0.22μm membrane and store at 2℃~8℃.

[0093] Will be prepared β 2 -The microsphere protease label complex (labeled HRP or AP) is added to the stability solution described above to prepare a working solution.

[0094] 2. Performance test of enzyme label complex:

[0095] 2.1. AP enzyme label complex test method:

[0096] (1) Use a pipette to accurately pipette 150μL of magnetic bead working solution to a 1.5mL centrifuge tube, and add 10μL of β to it in turn 2 -MG 0.12μg / mL sample or 19.5μg / mL sample, 30μL enzyme-labeled complex, then use a pipette to disperse the mixture evenly, and start timing. After 5 minutes, the centrifuge tube was placed on the magnetic stand, 30 seconds later, the clear liquid was aspirated, and ...

Embodiment 2

[0120] 1. C-reactive protease label complex preservation solution

[0121] 1L specific formula: MOPS 4.18g, NaCl 8.8g, CaCl 2 0.55g, aminopyrine 5g, casein 10g, gelatin 5g, trehalose 30g, glycerol 50mL, PEG20000, 0.5mL, Proclin300, 0.5mL. Prepared with sterile water, adjust the pH to 7.8, and store at 2-8°C after filtration with 0.22μm membrane

[0122] Add the prepared enzyme label complex (HRP or AP) to the stability solution described below to prepare a working solution.

[0123] 2. Performance test of enzyme label complex:

[0124] 2.1. AP enzyme label complex test method:

[0125] (1) Use a pipette to accurately pipette 150μL of magnetic bead working solution to a 1.5mL centrifuge tube, add 10μL of 1.15μg / mL CRP sample or 20.34μg / mL sample, 30μL of enzyme-labeled complex to it in turn, and then use the pipette Disperse the mixture evenly and start timing. After 5 minutes, the centrifuge tube was placed on the magnetic stand, 30 seconds later, the clear liquid was aspirated, and...

Embodiment 3

[0149] 1. NT-proBNP enzyme label complex preservation solution

[0150] 1L specific formula: Na 2 HPO 4 .12H 2 O 2g, KH 2 PO 4 0.2g, NaCl 8.8g, KCl 0.2g, aminopyrine 10g, casein 5g, trehalose 20g, glycerol 60mL, Tween 20, 0.5mL, Proclin300, 0.5mL. Prepared with sterile water, adjust the pH to 6.9, and store at 2-8°C after filtration with 0.22μm membrane

[0151] Add the prepared enzyme label complex (HRP or AP) to the stability solution described below to prepare a working solution.

[0152] 2. Performance test of enzyme label complex:

[0153] 2.1. AP enzyme label complex test method:

[0154] (1) Use a pipette to accurately pipette 150μL of the magnetic bead working solution to a 1.5mL centrifuge tube, add 10μL of 300.21pg / ml or 3000.08pg / mL NT-proBNP sample, 30μL of enzyme-labeled complex to it, and then pipette The gun will disperse the mixture evenly and start timing. After 5 minutes, the centrifuge tube was placed on the magnetic stand, 30 seconds later, the clear liquid was a...

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PUM

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Abstract

The invention relates to the technical field of chemiluminescence immune assay, in particular to a composition and an application of the composition as enzyme labeled compound preserving fluid. The composition can improve the stability of an enzyme labeled compound, prolong a storage life of the enzyme labeled compound and stably preserve various enzyme labeled compounds for a long term. Experiments show that the stability difference between the un-aged enzyme labeled compounds of beta 2-microglobulin, C reactive protein or NT-proBNP (N-terminal pro-brain natriuretic peptide) and the aged enzyme labeled compounds for 6 days is within plus-minus 10%.

Description

Technical field [0001] The invention relates to the technical field of chemiluminescence immunoassays, in particular to a composition and its application as an enzyme-labeled complex preservation solution. Background technique [0002] Chemiluminescence immunoassay is an analysis technique that combines highly sensitive chemiluminescence detection with highly specific immune response to detect various haptens, antigens, antibodies, enzymes, hormones, fatty acids, drugs and vitamins. Enzymatic chemiluminescence (also called indirect chemiluminescence) refers to the labeling of related enzymes on antibodies or antigens. The enzyme-labeled antibody-antigen complexes continuously emit visible light under the action of a luminescent substrate. Typical labels are alkaline phosphatase. (AP) and horseradish peroxidase (HRP), the corresponding marker substrates are luminol and its derivatives and 1,2-dioxane derivatives (AMPPD). Chemiluminescence products have been rapidly promoted in cl...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N21/76
CPCG01N33/535G01N21/76
Inventor 刘萌萌
Owner SINOCARE
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