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A metal ion-qlisa immunodetection signal amplification kit and preparation method thereof

A metal ion and immunological detection technology, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., to achieve the effect of increasing the binding rate and enhancing the sensitivity

Active Publication Date: 2021-01-22
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no research on the impact of metal ions on biological detection. In the physiological environment, there are more or less many metal ions. Coenzymes, hormones and other aspects play an important role, for example, Ca in human plasma 2+ and Mg 2+ The characteristic concentrations of are very close, being 1.16×10 -6 ~1.32×10 -6 mol / mL and 0.5×10 -6 ~0.65×10 -6 mol / mL

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  • A metal ion-qlisa immunodetection signal amplification kit and preparation method thereof
  • A metal ion-qlisa immunodetection signal amplification kit and preparation method thereof
  • A metal ion-qlisa immunodetection signal amplification kit and preparation method thereof

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preparation example Construction

[0030] In the present invention, the carboxyl-coated water-soluble quantum dots in the second reaction antibody labeled with carboxyl-coated water-soluble quantum dots include silica-coated quantum dots QDs@SiO 2 , QDs@PMAH coated with amphiphilic polymer or QDs@MPA modified with 3-mercaptopropionic acid. In the present invention, the quantum dots QDs@SiO coated with silicon dioxide 2 , the sources of quantum dots QDs@PMAH coated with amphiphilic polymers or quantum dots QDs@MPA modified with 3-mercaptopropionic acid are not particularly limited, and conventional commercially available products can be used or self-prepared. The present invention is illustrated by taking the coating method of silicon dioxide (silane coupling agent) and preparing surface carboxylated CdSe / ZnS quantum dots by the reverse microemulsion method as an example. The preparation method of the water-phase quantum dots of surface carboxylation based on the coating method (silane coupling agent) of silico...

Embodiment 1

[0059] Ca 2+ -QLISA (QDs@SiO 2 ) kit preparation and detection

[0060] (1) Composition of the kit

[0061] First reaction antibody for coating: CRP monoclonal antibody 1, purchased from Beijing Baichuan Feihong Biotechnology Co., Ltd., the product catalog number is 7D9, and the concentration is 2.2 mg / mL.

[0062] Second reaction antibody for conjugation: CRP monoclonal antibody 2, purchased from Hangzhou Qitai Co., Ltd., the product catalog number is MC02, and the concentration is 5 mg / mL.

[0063] Carboxyl-coated water-soluble quantum dot-labeled secondary reactive antibody: CaCl 2 Join QDs@SiO 2 Conjugated CRP mAb 2, CaCl 2 The concentration is 5×10 -6 mol / mL.

[0064] Standard product: CRP standard product was purchased from Hangzhou Qitai Co., Ltd., and was diluted to relevant concentration with diluent.

[0065] Washing solution: 1×10 containing 0.05% Tween 20 (volume ratio) -5 mol / mL phosphate buffer, pH 7.4.

[0066] Diluent: 1×10 containing 30% newborn calf...

Embodiment 2

[0083] Mg 2+ -QLISA (QDs@SiO 2 ) kit preparation and detection

[0084] First reaction antibody for coating: CRP monoclonal antibody 1, purchased from Beijing Baichuan Feihong Biotechnology Co., Ltd., the product catalog number is 7D9, and the concentration is 2.2 mg / mL.

[0085] Second reaction antibody for conjugation: CRP monoclonal antibody 2, purchased from Hangzhou Qitai Co., Ltd., the product catalog number is MC02, and the concentration is 5 mg / mL.

[0086] Carboxyl-coated water-soluble quantum dots labeled secondary reactive antibody: MgCl 2 Join QDs@SiO 2 Conjugated CRP mAb 2, MgCl 2 The concentration is 3×10 -5 mol / mL.

[0087] Standard product: CRP standard product was purchased from Hangzhou Qitai Co., Ltd., and was diluted to relevant concentration with diluent.

[0088] Washing solution: 1×10 containing 0.05% Tween 20 (volume ratio) -5 mol / mL phosphate buffer, pH 7.4.

[0089] Diluent: 1×10 containing 30% newborn calf serum (volume ratio) -5 mol / mL pho...

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Abstract

The invention provides a metal ion-QLISA immunodetection signal amplification kit and a preparation method thereof, belongs to the technical field of in-vitro diagnosis and detection, and the kit comprises the following components: a detection plate coated with a first reaction antibody, a carboxyl-coated water-soluble quantum dot-labeled second reaction antibody, a coupling antibody, a diluent, astandard solution and a washing solution, wherein the water-soluble quantum dot-labeled second reaction antibody s comprises divalent metal ions and trivalent metal ions. According to the invention,the detection fluorescence of the metal ion QLISA immunodetection signal amplification kit provided by the invention can be enhanced to 2.5-4 times, and the sensitivity can be enhanced to 2-4 times; the kit has the characteristics of wide detection range and stable and accurate detection result.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis and detection, and in particular relates to a metal ion-QLISA immunoassay signal amplification kit and a preparation method and application thereof. Background technique [0002] In recent years, because quantum dot materials have many excellent fluorescence characteristics such as high fluorescence efficiency, high stability, large Stockes shift, narrow and symmetrical emission spectrum, single wavelength and multicolor excitation, etc., the detection technology based on quantum dot materials has been obtained. With the great development, it is widely used in the qualitative and quantitative detection of toxic substances, excessive substances, inflammatory factors, cancer factors and drugs in food. The QLISA method has high sensitivity and can be used for quantitative detection. It has been widely used in the detection of related markers. For example, QLISA using QDs as fluorescent mar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/543
CPCG01N33/543G01N33/588G01N33/68
Inventor 吴瑞丽王盼盼李金洁吕雁冰申怀彬李林松
Owner HENAN UNIVERSITY