Latex-enhanced immunoturbidimetric assay kit for NGAL, and preparation method thereof

A detection kit, latex immunization technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of restricting the application of domestic reagents and low correlation of detection reagents, achieving good correlation, improved stability, and high sensitivity Effect

Inactive Publication Date: 2018-04-20
捷和泰(北京)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the correlation between the existing domestic self-produced NGAL reagents and the detection reagents produced by BIOPORTO company is low, which greatly limits the clinical application of domestic reagents

Method used

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  • Latex-enhanced immunoturbidimetric assay kit for NGAL, and preparation method thereof
  • Latex-enhanced immunoturbidimetric assay kit for NGAL, and preparation method thereof
  • Latex-enhanced immunoturbidimetric assay kit for NGAL, and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: The sensitization of the NGAL latex immunoturbidimetric assay kit using polyethylene glycol hexammonia as a blocking agent

[0050] 1. The preparation of NGAL latex immunonephelometry detection kit of the present invention

[0051] Reagent R1 includes: 25mM HEPES buffer (pH7.5), 150mM NaCl, 10mM EDTA-2Na, 1%wtBSA, 1%wt polyethylene glycol 6000, 0.01%wt Tween-20, 1%wt rabbit serum or Mouse serum, 0.1% wt sodium azide.

[0052] Reagent R2 includes: rabbit anti-human polyclonal antibody or mouse anti-human monoclonal antibody (5mg / mL), 10%wt latex particles (particle size 300nm, surface charge 0.15mmol / g), 0.5%wt polyethylene glycol hexaamine (polyethylene glycol molecular weight 2000 or 5000), 0.1% wt BSA, 0.1% wt sodium azide and 50 mM HEPES buffer (pH 7.5).

[0053] Calibrators include: 10mM HEPES buffer (pH7.5), 1% wtBSA, 0.1% wt Tween-20, 0.1% wt sodium azide and NGAL recombinant protein, wherein the concentrations of NGAL recombinant protein are 150ng...

Embodiment 2

[0065] Embodiment 2: Serum influence rate of the NGAL latex immunoturbidimetric assay kit using polyethylene glycol hexammonia as a blocking agent

[0066] The effects of six blocking agents with better reactivity (BSA, N101, PEG(5000)-6N, PEG(2000)-6N, PEG1000, mPEG-N-5000) on the accuracy of reagent measurement were compared. The specific method is: add human NGAL recombinant antigen with a concentration of 20,000 ng / mL to 0.9% NaCl, human-derived normal serum (EXA-N), and human-derived abnormal serum (EXA-A), respectively, so that the final concentration of NGAL in each matrix is 200ng / mL. The samples of these three matrices were tested using the above-mentioned reagents blocked by the blocking agent, and compared with the detection results of 0.9% NaCl.

[0067] Experimental results such as image 3 As shown, compared with the traditional blocking agent BSA, the serum influence rate of PEG(5000)-6N, PEG1000, mPEG-N-5000 blocking monoclonal antibody coating reagent is clo...

Embodiment 3

[0068] Example 3: The specificity of the NGAL latex immunoturbidimetric assay kit using polyethylene glycol hexammonia as a blocking agent

[0069] 1. The preparation of NGAL latex immunoturbidimetric assay kit (J&W) of the present invention

[0070] Reagent R1 includes: 50mM HEPES buffer (pH8.0), 150mM NaCl, 10mM EDTA-2Na, 1%wtBSA, 0.8%wt polyethylene glycol 6000, 0.01%wt Tween-20, 1%wt rabbit serum and 0.1% wt sodium azide.

[0071] Reagent R2 includes: rabbit anti-human polyclonal antibody (5mg / mL), 10%wt latex particles (particle diameter 200nm, surface charge 0.2mmol / g), 0.5%wt polyethylene glycol hexaamine (polyethylene glycol molecular weight is 5000), 0.1% wtBSA, 0.1% wt sodium azide and 50 mM HEPES buffer (pH 8.0).

[0072] Calibrators include: 10mM HEPES buffer (pH8.0), 1% wtBSA, 0.1% wt Tween-20, 0.1% wt sodium azide and NGAL recombinant protein, wherein the concentrations of NGAL recombinant protein are 150ng / mL, 600ng / mL, respectively mL, 1500ng / mL, 3000ng / mL, ...

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Abstract

The invention discloses a latex-enhanced immunoturbidimetric assay kit for neutropil gelatinase-associated lipocalin, and a preparation method thereof. Polyethylene glycol hexamine is selected as theblocking agent of an antibody binding latex particle. The kit used for latex-enhanced immunoturbidimetric assay of the NGAL has the advantages of high detection specificity, high sensitivity and goodstability, and can efficiently detect the NGAL content in urine, plasma and serum, and the detection result is well associated with fluorescence immunochromatography and enzyme linked immunosorbent assay.

Description

technical field [0001] The present invention relates to a kind of NGAL latex immune turbidimetric method detection kit and its preparation method, specifically, relate to a kind of NGAL latex immune turbidimetric method detection kit and its preparation method using polyethylene glycol hexammonia as blocking agent . Background technique [0002] Acute kidney injury (AKI) is a disease with high morbidity and mortality. Due to the unknown pathogenesis of the disease and the lack of accurate and reliable early diagnostic markers, effective therapeutic drugs have not yet appeared clinically. Traditional markers of renal injury (such as serum creatinine, cystatin C, α1 microglobulin, β2 microglobulin, and retinol-binding protein) are mostly functional markers, but most of the structural changes in AKI occur in functional markers. Therefore, structural kidney injury markers are more sensitive for the diagnosis of AKI, among which the most well-studied and the most significant cha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577
CPCG01N33/577G01N33/6893G01N2800/34
Inventor 周淼
Owner 捷和泰(北京)生物科技有限公司
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