a protein chip

A protein chip, chip technology, applied in instruments, biological material analysis, measurement devices, etc., can solve the problems of undetectable technology, false negative results, difficulty in diagnosing cancer, etc., and achieve the effect of improving sensitivity and reducing preparation costs

Active Publication Date: 2019-01-11
天津东亚生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, it has been confirmed by clinical testing that the content of HLA-G in the serum of patients with advanced tumors does not increase with the increase of tumor cells. Undetectable, producing false negative results
This brings serious difficulties to the clinical application of HLA-G in the diagnosis of cancer
[0007] We also learned that certain compounds can interfere, causing false positives in the assay

Method used

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Experimental program
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Effect test

Embodiment 2

[0039] Preparation of various polymer plates with immunocompetent wells

[0040] 1. Prepare small hole arrays on various plates: use hot pressing or drilling to form holes, the diameter of the holes is 3-6mm small holes, the volume is preferably 10-50 microliters, and the hole distance depends on the measuring instrument. Suitable microtiter plates can also be used.

[0041] 2. Coating primary antibody: Add 10-50 microliters of appropriately diluted primary antibody to each well at pH 7.0-9.5, incubate overnight at 4°C or 2 hours at 37°C, and adsorb the primary antibody on the plate On the wall of the small hole, suck out the antibody solution after the reaction.

[0042] 3. Blocking: Add 10-50 microliters of blocking solution to each well, incubate at 37°C for 2 hours, or overnight at 4°C. After the reaction, the blocking solution was shaken off, air-dried and sealed at 4°C for future use.

[0043] Of course, the immunological microassay plate is used to prepare the polyme...

Embodiment 3

[0045] Commonly used polymer balls include polypropylene balls, nylon balls, polystyrene balls and their microspheres. The diameter of the high molecular polymer ball is between 4-6mm, and the diameter of the microsphere is between 10-60nm. Some of the balls have a smooth surface and some have a rough surface.

[0046] (1) Polypropylene beads or microspheres The primary antibody can be coated on the surface of the beads or microspheres by electrostatic adsorption, see Example 2 for details.

[0047] (2) React the nylon ball with 1-4 mol / L hydrochloric acid for 1-6 hours at room temperature to expose the amino and carboxyl groups, and then coat the primary antibody on the surface of the nylon ball with carbodiimide or glutaraldehyde .

[0048] (3) Polystyrene beads or microspheres coated with the first antibody The first antibody can be coated on the surface of polystyrene beads or microspheres by electrostatic adsorption, see Example 2; polystyrene beads Balls or microspher...

Embodiment 4

[0050] Using the glass protein chip and polymer chip prepared in embodiment 1 and embodiment 2 to measure HLA-G

[0051] 1. Determination of the serum to be tested: add the No. 1 serum to be tested to each of the first vertical row of the glass slide (or ball) or the polymer plate after binding the first antibody and blocking the glass chip or polymer chip. In the active wells, serum No. 2 is added to each active well in the second vertical row, and so on, serum No. 3 to serum n are added to each active well in the third to nth vertical rows in turn, and each active well is added 10-50 microliters of serum to be tested.

[0052] 2. Incubate the active glass slide (or ball) or polymer plate (or ball) at 37°C for 60 minutes, shake once (5 seconds) in the middle of the reaction, suck out the serum, and discard it after disinfection.

[0053] 3. Washing: Add 10-50 microliters of washing solution to each active well, wash 5-8 times, shake for 5 seconds each time, suck out the wash...

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Abstract

The invention discloses a novel protein chip. A substrate material of the novel protein chip adopts a glass sheet with holes, a polymer plate with holes, glass balls or polymer balls which are represented by an immune microdetermination plate to replace a substrate material NC (Nitrocellulose) film of a traditional protein chip. A general immunity analyzer replaces an interpretoscope of the traditional protein chip by detecting color or light intensity generated by an immunoreaction. The novel protein chip and immune software are matched to jointly qualitatively and quantitatively detect multiple immune ligands in a solution. The novel protein chip for broad spectrum tumors is an instrument for jointly qualitatively and quantitatively detecting cancers by using a plurality of tumor markers. According to the novel protein chip for the broad spectrum tumors, false positive and false negative can be effectively eliminated, the accuracy of detection is improved, and the manufacturing cost of the chip is reduced.

Description

technical field [0001] The invention relates to the technical fields of molecular biology, biomedicine and clinical immunoassay, and in particular to a novel protein chip. Background technique [0002] Tumor is a common disease and frequently-occurring disease, and its incidence rate ranks second. According to the report of the World Health Organization in 1997, the annual death toll of cancer patients in the world is as high as 6.6 million, and by 2020, the number of cancer patients in the world will reach 15 million. Various malignant tumors seriously threaten human health. In the early stage of tumor onset, there are often no subjective symptoms, and once the subjective symptoms are found, it has reached the middle and late stage that cannot be treated. Today, when the pathogenesis and treatment of tumors have not been completely resolved, in order to reduce the mortality of cancer patients, it is necessary to carry out basic-level tumor census work, so as to achieve ea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/552G01N33/531G01N33/532G01N21/76
CPCG01N21/76G01N33/531G01N33/532G01N33/552G01N33/57484G01N2800/00
Inventor 苏殿杰
Owner 天津东亚生物技术有限公司
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