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Anti-biotin interference chemiluminiscence immunoassay kit and application thereof

A chemiluminescence immunoassay, chemiluminescence reagent technology, applied in the direction of chemical reaction of materials for analysis, chemiluminescence/bioluminescence, biological testing, etc., can solve problems such as biotin interference, achieve low performance impact, eliminate free biological the effect of the interference of the hormone, avoiding false positive and/or false negative results

Pending Publication Date: 2020-05-08
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there is no method to solve the problem of biotin interference well in the avidin-biotin system

Method used

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  • Anti-biotin interference chemiluminiscence immunoassay kit and application thereof
  • Anti-biotin interference chemiluminiscence immunoassay kit and application thereof
  • Anti-biotin interference chemiluminiscence immunoassay kit and application thereof

Examples

Experimental program
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preparation example Construction

[0109] In some embodiments of the present invention, the preparation method of the anti-interference agent includes: step S1, contacting the carrier with the active molecule; preferably, the contacting is performed in the first buffer system. In some embodiments of the above method, the pH value of the first buffer system is 7.0 to 9, preferably 7.1 to 8.0, more preferably 7.2 to 7.8, more preferably 7.3 to 7.7, further preferably 7.35 to 7.50, most preferably 7.40 . In this embodiment, examples of the pH value of the first buffer solution system include 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 9.0 and the like.

[0110] In some embodiments of the above method, the pH value of the first buffer system is 7.0 to 9, preferably 7.1 to 8.0, more preferably 7.2 to 7.8, more preferably 7.3 to 7.7, further preferably 7.35 to 7.50, most preferably 7.40 . In this embodiment, examples of the pH value of the first buffer solution system include 7.0, 7.1, 7.2, 7.3, 7.4, 7.5...

Embodiment 1

[0143] In the first step, take 10 mg of carboxyl-functionalized silicon-based microspheres (particle size 15 nm, pore size 2 nm) in a 2 mL centrifuge tube, add 0.02 M PBS (pH 7.4) buffer solution, centrifuge at 10,000 rpm at 4 ° C, and wash once for 15 min.

[0144] In the second step, add 200uL PBS buffer solution to ultrasonically disperse evenly, then add 150uL 10mg / mL SA aqueous solution, supplement PBS buffer solution to the microsphere reaction concentration of 20mg / mL, and stir overnight at room temperature.

[0145] In the third step, the SA microspheres were centrifuged with 0.02M PBS (pH 7.4) buffer solution containing 0.5% Tween-20, centrifuged at 10,000 rpm at 4°C for 15 min, washed three times to remove unadsorbed SA, and finally washed with 0.02M PBS (pH 7.4). 7.4) Dilute the buffer solution to 10 mg / mL.

Embodiment 2-7

[0147]The preparation method is the same as that of Example 1, except that the carboxyl-functionalized silicon-based microspheres with different particle diameters and / or pore diameters are used in each embodiment (see Table 1).

[0148] Preparation of anti-interference agent of the present invention in covalent coupling mode

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Abstract

The invention relates to an anti-biotin interference chemiluminiscence immunoassay kit and application thereof in the technical field of chemiluminiscence immunoassay. The kit comprises: a) an anti-interference agent, which comprises a carrier and an active molecule, wherein the carrier is a porous medium, the active molecule is filled in the carrier and can specifically bind with biotin molecules; b) an immune molecule, which can immunologically bind with a to-be-detected target molecule; and c) a chemiluminescent reagent. The kit provided by the invention comprises an anti-interference agent, according to the anti-interference agent, an active molecule which can be specifically combined with biotin molecules is used as guest molecule and is filled in a carrier in a proper mode to form amesoporous assembly host-guest system, so that the kit avoids false positive or false negative results caused by free biotin.

Description

technical field [0001] The invention belongs to the technical field of chemiluminescence immunoassay, and in particular relates to a biotin-resistant chemiluminescence immunoassay kit and an application thereof. Background technique [0002] Biotin-avidin system (Biotin-Avidin-System, BAS) is a new biological reaction amplification system developed in the late 1970s. The BAS system has the advantages of high affinity, high sensitivity, high stability and other advantages of signal amplification labeling technology. By combining the two, macromolecular bioactive substances such as antigens and antibodies can be coupled. Their combination is rapid, specific, stable, and has a multi-stage amplification effect. At present, the BAS system is mainly used in immunology, molecular biology and other fields. In the practical application of in vitro diagnosis, it has great advantages. The biggest disadvantage of this method is the interference of biotin, which causes detection error...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N21/76
CPCG01N33/5306G01N21/76
Inventor 章春奇金鑫赵卫国刘宇卉李临其他发明人请求不公开姓名
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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