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Time-resolved immunochromatography test strip and kit for detecting S100B protein as well as preparation method and application of time-resolved immunochromatography test strip

An immunochromatographic test paper and time-resolving technology, which is applied in the biological field, can solve the problems of the stable period of the kit and the short half-life of the nuclide, the high price of the fluorescent immunoassay, and the complicated operation, so as to reduce the process of preparing the binding pad and improve the The effect of sensitivity and simple preparation process

Active Publication Date: 2021-11-30
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among these methods, the price of fluorescent immunoassay is relatively high, and it is difficult to popularize. Both immunoradiometric assay and radioimmunoassay have certain radioactivity, and the stability period of the kit and the half-life of the nuclide are relatively short. Restricted by complex operation

Method used

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  • Time-resolved immunochromatography test strip and kit for detecting S100B protein as well as preparation method and application of time-resolved immunochromatography test strip
  • Time-resolved immunochromatography test strip and kit for detecting S100B protein as well as preparation method and application of time-resolved immunochromatography test strip
  • Time-resolved immunochromatography test strip and kit for detecting S100B protein as well as preparation method and application of time-resolved immunochromatography test strip

Examples

Experimental program
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Effect test

preparation example Construction

[0042] The present invention also provides a preparation method of a time-resolved immunochromatographic test strip, the preparation method comprising:

[0043] Coating membrane processing step: respectively immobilizing the S100B monoclonal capture antibody and goat anti-rabbit IgG antibody recognizing a single epitope on the coating membrane to form a detection area and a control area;

[0044] Sample pad processing steps: prepare fluorescent microsphere-labeled S100B polyclonal detection antibody and fluorescent microsphere-labeled rabbit IgG, and spray on the sample pad;

[0045] Assembly steps: Lap and paste the sample pad, coating film and absorbent paper on the backing in sequence.

[0046] As a further preferred solution, in the sample pad processing step of the present invention, the method for preparing the S100B polyclonal detection antibody labeled with fluorescent microspheres is as follows:

[0047] (1) Dialyze the S100B polyclonal detection antibody overnight, ...

Embodiment 1

[0054] Example 1: Screening of S100B Antibody

[0055] In this example, conventional serum experiments were used to select monoclonal capture antibodies and The polyclonal labeled antibodies were paired for experiments, as shown in Table 1 below. In the experiment, 12 groups of paired experiments were set up, and each group was tested with 10 S100B negative and positive serum samples. The composition of the 12 groups of paired experiments is shown in Table 2. For the paired results, see Figure 1-12 .

[0056] Table 1: Antibody Pairing Experiment Table

[0057]

[0058] The sequence of AB2 is SEQ ID NO: 1, the sequence of AB3 is SEQ ID NO: 2, AB1 is the antibody with the article number V01003 produced by Nanjing GenScript Biotechnology Co., Ltd., and AB4 is the antibody produced by Nanjing GenScript Biotechnology Co., Ltd. The antibody produced by the company is V01001.

[0059] SEQ ID NO: 1: MSELEKAMVALIDVFHQYSGREGDKHKLKKSELKELINNELSHFLEEIKEQEVVDKVMET

[0060] SEQ ID ...

Embodiment 2

[0068]A time-resolved immunochromatographic test strip for detecting S100B, comprising a substrate, a sample pad, a coating film and absorbent paper sequentially arranged on the substrate, the sample pad is coated with S100B marked with fluorescent microspheres Polyclonal labeled antibody (SEQ ID NO: 1, derived from Raybiotech, CAT#144-00676) and rabbit IgG labeled with fluorescent microspheres (Hangzhou Boyin Biotechnology Co., Ltd., Cat. No.: N160701), the coated membrane includes parallel settings , and a detection area and a control area at a distance of 0.5 cm from each other, the detection area is close to the absorbent paper, the control area is close to the sample pad, and the detection area is coated with S100B polyclonal capture that recognizes a single antigenic epitope Antibody (SEQ ID NO: 2, derived from Ruiboao (Guangzhou) Biotechnology Co., Ltd.), and the control area was coated with goat anti-rabbit IgG antibody (Hangzhou Boyin Biotechnology Co., Ltd., catalog n...

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Abstract

The invention discloses a time-resolved immunochromatography test strip for detecting S100B protein, wherein the time-resolved immunochromatography test strip comprises a bottom lining, and a sample pad, a coating film and water-absorbent paper which are sequentially arranged on the bottom lining; the sample pad is coated with a fluorescent microsphere-labeled S100B polyclonal detection antibody and a fluorescent microsphere-labeled rabbit IgG; the coating film comprises a detection area and a control area which are arranged in parallel; a rear spacer region is arranged between the detection area and the control area, the detection area is coated with an S100B monoclonal capture antibody for recognizing a single epitope, and the control area is coated with a goat anti-rabbit IgG antibody; and the coating film is a nitrocellulose film combined with a light-transmitting polymer. The invention also discloses a preparation method of the time-resolved immunochromatographic test strip and a kit. The time-resolved fluorescent microspheres of lanthanum are used for marking the antibody, and the concentration of the S100B protein in a sample can be detected by combining an immunochromatography technology and adopting a double-antibody sandwich method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a time-resolved immunochromatographic test strip for detecting S100B protein, a kit, a preparation method and application thereof. Background technique [0002] The central nervous system-specific protein (S100B) is named for its ability to dissolve 100% in saturated ammonium sulfate solution. At present, there are 20 members of this family that have been discovered, most of which exist in the form of homodimers. S100B is an acidic calcium-binding protein with a molecular weight of 21KD. Structurally, S100B contains two EF chiral calcium-binding regions and an auxiliary Zn 2+ Binding site, via Zn 2+ The combination of histidine residues in the binding site changes the structure of S100B, shortens the distance between the calcium binding sites, and plays the role of calcium sensor protein. S100B mainly functions as an intracellular calcium receptor protein in the living body, and as...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/558G01N33/543G01N33/535G01N21/64
CPCG01N33/68G01N33/558G01N33/54306G01N33/54313G01N33/535G01N21/6408
Inventor 徐部灼宋旭东唐浩黄若磐
Owner RAYBIOTECH INC GUANGZHOU
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