Immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and preparation method thereof

A technology of lipocalin and neutrophils, applied in the field of clinical laboratory medicine, can solve the problems of limiting clinical application potential and inability to distinguish and measure different molecular forms of HNL

Inactive Publication Date: 2016-08-31
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Chinese patent (ZL201220497401) discloses an up-conversion luminescence rapid quantitative device for HNL detection. This technology partially overcomes the shortcomings of current HNL detection technology but cannot distinguish between different molecular forms of HNL, thus limiting the clinical application potential of this method
At present, there is no technology disclosure of combined simultaneous quantitative and rapid detection of HNL

Method used

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  • Immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and preparation method thereof
  • Immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and preparation method thereof
  • Immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1: Composition and preservation of an immunochromatographic device for simultaneous quantitative detection of different molecular forms of human neutrophil lipocalin (HNL)

[0071] 1) The composition of the device: 1 bottle of sample diluent freeze-dried powder, several UCP immunochromatography test strips, and 1 set of standard solution freeze-dried powder;

[0072] 2) Storage conditions of the device: storage at room temperature.

Embodiment 2

[0073] Example 2: Preparation of a UPT immunochromatography test strip for simultaneous quantitative detection of different molecular forms of human neutrophil lipocalin (HNL) immunochromatography device

[0074] 1) Solution preparation

[0075] a) Preparation of fMLP solution

[0076] Firstly, fMLP (purchased from Sigma) was dissolved in dimethyl sulfoxide (DMSO) to prepare a 50 mM stock solution, and then the 50 mM fMLP stock solution was diluted into a 5 μmol / L fMLP solution with 50 mM HEPES solution at pH=7.4.

[0077] b) Preparation of heparin sodium solution

[0078] Sodium heparin (purchased from Sigma) was dissolved in 50 mM HEPES (pH 7.4) solution to prepare a 14 U / mL sodium heparin solution.

[0079] c) Antibody solution preparation

[0080] Anti-MMP-9 polyclonal antibody (purchased from abcam company, ab5707), anti-three molecular forms of human HNL monoclonal antibody (monoclonal antibody prepared by immunizing mice with human homodimer HNL as antigen, can bind ...

Embodiment 3

[0116] Embodiment 3: A kind of standard curve of simultaneous quantitative detection different molecular form human neutrophil lipocalin (HNL) immunochromatography device

[0117] 1) Solution preparation

[0118] a) Preparation of sample diluent

[0119] The sample diluent is obtained by diluting the sample diluent lyophilized powder.

[0120] Sample diluent lyophilized powder is 20mL, pH 7.2 containing 100mmol / L HEPES solution of 200mmol / L NaCl, 0.5% (vol / vol) Tween-20, 1% (wt / vol) BSA. μmol / L filter to filter and freeze-dry. The sample diluent is obtained by adding 20 mL of deionized water to the lyophilized powder of the above sample diluent and mixing.

[0121] b) Preparation of HNL standard concentration gradient solution

[0122] The mother solution of concentration gradient solution of HNL standard substance is obtained by adding deionized water to standard substance freeze-dried powder. The standard freeze-dried powder contains 400 μg / mL monomer, homodimer, or het...

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Abstract

The invention discloses an immunochromatography device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and a preparation method thereof, and belongs to the field of laboratory medicine. The device is composed of sample diluent freeze-dried powder, standard substance freeze-dried powder and an immunochromatography test strip. The immunochromatography test strip is composed of a liner plate, two blood filtration membranes, a sample flow pad, a UCP combination pad, an analysis membrane and an absorption pad. The UCP combination pad is coated with an antibody capable of resisting three molecular forms of HNL with UCP markers; the analysis membrane is provided with three parallel detection regions (T1, T2 and T3) and a quality control region C, and the three detection regions are coated with an antibody capable of resisting a matrix metal protease-9, an antibody capable of resisting a monomer HNL and an antibody capable of resisting homodimer; the quality control region C is coated with an antibody from animal immune globulin with the UCP marker. One sample can detect three molecular forms of HNL in a combined mode on the same test strip, and the device has the advantages of being rapid, quantitative, easy and convenient to use and the like and has the large scientific research and clinic application prospects.

Description

technical field [0001] The invention belongs to the technical field of clinical laboratory medicine, and in particular relates to an immunochromatographic device for synchronously and quantitatively detecting different molecular forms of human neutrophil lipocalin (HNL) and a preparation method thereof. Background technique [0002] Human neutrophil lipocalin (human neutrophil lipocalin, HNL) was originally a glycoprotein discovered in neutrophil granules by Xu S. et al. (Scand.J.Clin.Lab Invest 54,365-376) , a member of the lipocalin superfamily, has a similar spatial structure to other lipocalins. The protein is also called neutrophil gelatinase associated lipocalin (neutrophil gelatinase associated lipocalin, NGAL), also known as human lipocalin 2 (lipocalin 2) and so on. Cai Linjun et al. (Clin.J.Am.Soc.Nephrol.2010,5:2229-35) found that urinary HNL exists in various molecular forms, including monomer (about 25kDa), homodimer (about 45kDa) and A heterodimer (about 135k...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/543
CPCG01N33/543G01N33/558
Inventor 蔡林君杨津陈雷赵冰姜春来孔维
Owner JILIN UNIV
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