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Protein-free coated plate confining liquid

A technology of coating plate and sealing liquid, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effects of short sealing time, stable properties and simple operation

Active Publication Date: 2014-03-05
BIOBASE BIODUSTRY (SHANDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Commonly used substances in the blocking solution include BSA, skim milk, complexin, etc., but most of the BSA contains antibody residues, which will lead to cross-contamination between antigens / antibodies. A high background is generated, which increases the background level; due to the complex composition of milk powder, and the sealed carrier is not easy to store for a long time, it is rarely used in the preparation of the kit; although the complex protein is relatively pure and stable, but due to The non-specific adsorption of ionic groups on the protein will increase the absorbance of the blank of the microwell plate after sealing, which will affect the linearity and sensitivity of the kit detection; at the same time, the coated plate after sealing treatment with several common blocking solutions should be kept at 2~8℃. If it is stored in the refrigerator for one week or destroyed at 37°C for one day, its biological activity will be lost by more than 50%.
[0007] It can be seen that after the application of conventional blocking solution, the background of the coated plate is relatively high, and the stability of the coated plate after sealing is poor

Method used

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  • Protein-free coated plate confining liquid
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  • Protein-free coated plate confining liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] The procedure of the hepatitis C core antigen detection kit (ELISA) established with the hepatitis C core antigen (HCV) monoclonal antibody coated in the embodiment of the present invention is the same as that of the comparative example 1, the difference is that in step 1, the amount of blocking solution coated on the plate is The components are:

[0065] Tris-HCl buffer solution (pH=7.4) 0.1mol / L,

[0066] PEG-20000 1g / L,

[0067] Tween-20 (Tween-20) 5g / L,

[0068] Trehalose 10g / L,

[0069] Sodium azide 1g / L.

[0070] Discard the coating solution in the coated plate, pat the coated plate dry, add the above-prepared coated plate blocking solution at 100 μL to 300 μL / well, and coat at 37°C for 2 hours; the rest of the operations are carried out as compared The operation is the same as in example 1. The test results are shown in Table 2 below.

[0071] Table 2 The OD values ​​of standard substances with different concentrations detected under different storage condi...

Embodiment 3

[0091] Table 4 Comparative Example 3 Detection Results

[0092]

[0093] Through the experimental data, it can be seen that the coated plates prepared in Example 1 and Comparative Examples 1, 2, and 3: in the absence of BSA blocking, the reagent blank absorbance will be relatively low; In the case of sugar, the prepared coated plate can be stored stably at 37°C for 60 days without damage, and the correlation coefficient is still above 0.99. If macromolecular substances or trehalose are added alone, the above requirements cannot be met.

Embodiment 2

[0095] The steps of the hepatitis C core antigen detection kit (ELISA) established with the hepatitis C core antigen (HCV) monoclonal antibody coated in the embodiment of the present invention are the same as in Example 1, except that the composition of the blocking solution for the coated plate in step 1 Divided into:

[0096] Tris-HCl buffer solution (pH=7.4) 0.01mol / L,

[0097] Polyvinylpyrrolidone 0.5g / L,

[0098] Tween-20 (Tween-20) 0.5g / L,

[0099] Trehalose 1g / L,

[0100] Sodium azide 0.1g / L.

[0101] The experimental operation flow is the same as that described in Example 1.

[0102] Example 3

[0103] The steps of the hepatitis C core antigen detection kit (ELISA) established with the hepatitis C core antigen (HCV) monoclonal antibody coated in the embodiment of the present invention are the same as in Example 1, except that the composition of the blocking solution for the coated plate in step 1 Divided into:

[0104] Tris-HCl buffer solution (pH=7.4) 0.05mol / L...

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Abstract

The invention belongs to the technical field of clinical in-vitro detection and particularly relates to protein-free coated plate confining liquid. The protein-free coated plate confining liquid contains Tris-HCL buffer liquor with PH of 7.4, a macromolecular substance, a surfactant, mycose and sodium azide, wherein the macromolecular substance is one or a mixture of two of PEG (Polyethylene Glycol)-20000 and polyvinyl pyrrolidone. The coated plate confining liquid disclosed by the invention not only can be combined with a blank position on the surface of the coated plate to achieve a confining effect for the coated plate, but also can keep biological activity of antigen / antibody on the surface of the coated plate longer after the coated plate leaves a low-temperature environment. The confining liquid provided by the invention is protein-free confining liquid, low in cost, simple to operate, stable in properties, shorter in sealing time in comparison with BSA (Bull Serum Albumin), and capable of achieving the effects of lowering cost and stabilizing the coated plate.

Description

technical field [0001] The invention belongs to the technical field of clinical in vitro detection, in particular to a protein-free coated plate blocking solution. Background technique [0002] In 1971, Engvall and Perlmann published an article on the use of enzyme-linked immunosorbent assay (enzyme linked immunosorbent assay, elisA) for the quantitative determination of IgG, which made the enzyme-labeled antibody technology for antigen localization developed in 1966 into the detection of trace substances in liquid specimens. test methods. The basic principle of this method is: ① Make the antigen or antibody bind to the surface of a certain solid phase carrier and maintain its immunological activity. ②The antigen or antibody is connected with an enzyme to form an enzyme-labeled antigen or antibody, which not only retains its immune activity, but also retains the activity of the enzyme. During the determination, the test specimen (determining the antibody or antigen in it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/54393G01N33/5767
Inventor 谭柏清王进甘宜梧
Owner BIOBASE BIODUSTRY (SHANDONG) CO LTD
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