Method for determining minim proteins based on magnetic pearl and nano gold probe

A nano-gold probe and assay method technology, which is applied in the field of protein assay, can solve the problems of failing to meet the clinical detection requirements, lower detection sensitivity, and complicated operating procedures, and achieve the effects of high sensitivity, convenient operation, and fewer operating steps

Inactive Publication Date: 2008-09-03
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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Problems solved by technology

However, the operation procedure of ELISA method is complicated, there are many steps, especially the time is long, it takes 5 or 6 hours, which brings some inconvenie

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  • Method for determining minim proteins based on magnetic pearl and nano gold probe
  • Method for determining minim proteins based on magnetic pearl and nano gold probe

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Embodiment Construction

[0035] Using this technology to analyze human p53 positive serum, the p53 protein content is 160pg / ml.

[0036] 1P53 Monoclonal Antibody Labeled Magnetic Beads

[0037] 1.1 Activate magnetic beads: Wash 300 μl magnetic beads (equivalent to 3 mg magnetic beads) twice with 300 μl 25mM MES (pH6), add 50 μl of 50 mg / ml EDC solution and 50 μl of 50 mg / ml NHS solution in ice bath, and incubate at room temperature for 30 minutes; Wash twice with 300 μl 25 mM MES (pH 6).

[0038]1.2 Labeling: Add 60 μl of p53 antibody, add 40 μl of 25mM MES (pH6), to make the final volume 100 μl; place at 4°C for 2 hours; wash 4 times with 300 μl of PBS (containing 0.1% BSA and 0.01% Tween-20), and then Resuspend the magnetic beads with 300 μl washing solution and store at 2-8°C for future use.

[0039] 2p53 Polyclonal Antibody and DNA Probe Labeling Nanogold

[0040] 2.1 Determine the optimal amount of antibody. Use 0.1M K2CO3 solution to adjust the pH value of the nano-gold solution to 9, and tak...

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Abstract

The invention relates to a high-sensitivity trace of protein measuring method based on bead and nm metal probe, comprising: labelling the bead using monoclonal antibody of protein to measure; labelling the polyclonal antibody of protein to measure on nm metal probe and at the same time labelling a DNA probe with biotin label on the nm metal probe; labelling horseradish peroxidase on the DNA probe of nm metal probe by biotin-streptavidin reaction to form nm metal probe; mixing the labelled bead and nm metal probe and protein sample and incubating for a period of time at 37 degree; and then washing the nm metal probes which do not react; developing using TMB development system, therefore the protein can be quantificationally detected. The detection time can be reduced to 1-1.5 hours and the sensitivity can be pg/ml.

Description

technical field [0001] The invention relates to a protein assay, in particular to a highly sensitive trace protein assay method based on magnetic beads and nano-gold probes, which can be applied to the medical field. Background technique [0002] Enzyme-linked immunoassay (ELISA) has been widely used in clinical laboratories, and can be used to detect various proteins such as various tumor markers, cytokines, and viral markers. There are also a variety of commercially available ELISA kits. However, the operation procedure of ELISA method is complicated, there are many steps, especially the time is long, it takes 5 or 6 hours, which brings some inconvenience to clinical application. If the detection time is shortened, the sensitivity of the detection will be reduced, and the detection requirements for clinical needs will not be met. Therefore, in order to meet the needs of rapid clinical detection, the development of rapid and highly sensitive protein detection methods has b...

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/532
Inventor 贾春平赵建龙金庆辉
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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