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Nanometer gold cage compound, and applications thereof in preparation of kit used for detecting cMyBP-C in human urine or blood

A nano-gold and composite technology, applied in the field of biotechnology applications, can solve the problems of lack of sensitivity and specificity, and achieve the effect of simple operation, high sensitivity and specificity, and easy observation

Active Publication Date: 2018-11-30
南京博天科智生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, other myocardial injury biomarkers such as myoglobin and CK-MB lack sensitivity or specificity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: the preparation of nano-gold cage complex

[0030] 1) Preparation of nano-gold cage solution: prepare HAuCl with deionized water 4 solution at room temperature using a syringe pump in HAuCl 4 Add citric acid dropwise to the solution, and then continue to stir until the color turns red and centrifuge to remove the supernatant to obtain cube hollow nanocrystals of different sizes, then resuspend the cube hollow nanocrystals with deionized water, and centrifuge to remove the supernatant. Much Au 3+ ionized, deionized water to resuspend the precipitate, and add 5mg / ml PVP monomer and 0.1% Tween to obtain a nano-gold cage solution; the dimensions of the cube hollow nano-crystals are 16nm and 42nm.

[0031] 2) Use 0.1M K 2 CO 3 Adjust the pH of the nano-gold cage solution to 8.5-9.0; react the specific single-stranded DNA with the nano-gold cage solution for 6-8 hours, add PVP monomer to make the final concentration 0.2%, adjust the pH to 7.4 Obtain the gol...

Embodiment 2

[0033] Embodiment 2: the preparation of nano-gold cage complex

[0034] It is basically the same as Example 1, except that the concentration of PVP monomer in step 2) is 20 mg / ml, and the final concentration of PVP monomer in step 3) is 1.0%.

Embodiment 3

[0035] The preparation of embodiment 3 nano-gold cage complexes

[0036]It is basically the same as in Example 1, except that the PVP monomer in step 2) is added at a concentration of 12 mg / ml, and the PVP monomer in step 3) has a final concentration of 0.6%.

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Abstract

The invention discloses a nanometer gold cage compound, and applications thereof in preparation of a kit used for detecting cMyBP-C in human urine or blood. The nanometer gold cage compound is capableof realizing covalent boding with a pair of anti-human cardiac myosin C monoclonal antibody BT09 and BT10, and forming the kit used for rapid detection of cMyBP-C in human urine or blood. The nanometer gold cage compound is used for replacing conventional spherical nanometer particles commonly used in colloidal gold test strips, is extremely high in human blood and / or urea cMyBP-C detection sensitivity and specificity; distinguishing light signals better that those of the spherical nanometer particles are generated based on the white background. At specific application conditions (size and diameter), the cube hollow nanometer crystal is capable of realizing covalent bond combination with DNA and proteins, biotin labelled single chain DNA is capable of connecting nanometer antibodies of two sizes, the detection signal is increased by 5000 to 10000 times of that of colloidal gold nanometer sphere POCT product. It is observed in results that the sensitivity is as high as 100fg / mL, reading of results is realized in 15min, and excellent stability and repeatability are achieved.

Description

technical field [0001] The invention belongs to the field of biotechnology applications, in particular to a nano-gold cage complex and its preparation method and application, in particular to the use of the nano-gold cage complex in the preparation of blood and / or urine cardiac myosin-binding protein C detection test paper Articles of application. Background technique [0002] Acute myocardial infarction (acute myocardial infarction, AMI) and / or injury (myocardial injury, MCI) is a class of clinically significant diseases with a high incidence. The mortality rate, hospitalization and medical treatment expenses of AMI and MCI are among the top in Europe, America, China and even the global population. Cardiac troponin (cTn) T / I, a biomarker of myocardial necrosis, is currently the preferred marker for diagnosis of AMI and risk stratification (Nat Rev Cardiol. 2012 Nov; 9(11):620-633). However, it is worth noting that the blood cTn concentration begins to increase 6-8 hours a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/558G01N33/68G01N33/577G01N33/58B82Y30/00B82Y15/00
CPCB82Y15/00B82Y30/00G01N33/532G01N33/558G01N33/577G01N33/587G01N33/68G01N2800/325
Inventor 张寄南张雷
Owner 南京博天科智生物技术有限公司
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