Novel protein chip

A protein chip, a new type of technology, applied in measurement devices, instruments, disease diagnosis, etc., can solve the problems of undetectable technology, false negative results, interference, etc., and achieve the effect of improving sensitivity and reducing preparation costs.

Active Publication Date: 2016-06-08
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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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0039] Preparation of various polymer plates with immunologically active holes

[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, and the volume is 10-50 microliters. The hole distance is determined by the measuring instrument. A suitable microtiter plate can also be used.

[0041] 2. Coating the primary antibody: Under the condition of pH 7.0-9.5, add 10-50 microliters of appropriately diluted primary antibody to each well, and incubate overnight at 4°C or 2 hours at 37°C to adsorb the primary antibody on the plate On the wall of the small hole, aspirate 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 4°C overnight. After the reaction, the blocking solution was thrown out, dried and sealed at 4°C for later use.

[0043] Of course, the immunoassay plate is used to prepare an immunologically act...

Embodiment 3

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

[0046] (1) Polypropylene beads or microspheres can be electrostatically adsorbed to coat the first antibody on the surface of the beads or microspheres, see Example 2 for details.

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

[0048] (3) Polystyrene beads or microspheres coated with the first antibody can be electrostatically adsorbed to coat the first antibody on the surface of the polystyrene beads or microspheres, see Example 2; polystyrene beads The spheres...

Embodiment 4

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

[0051] 1. Determination of the serum to be tested: Combine the first antibody and seal a good glass chip or polymer chip, and add the serum No. 1 to each of the first vertical row of the glass sheet (or ball) or polymer plate. In the active wells, serum No. 2 is added to each active hole in the second vertical row, and so on, serum No. 3 to No. n is added to each active hole in the third to nth vertical row in turn, and each active hole is added 10-50 microliters of serum to be tested.

[0052] 2. Incubate the active glass sheet (or ball) or polymer plate (or ball) at 37°C for 60 minutes, shake once during the reaction (5 seconds), aspirate the serum, and discard after disinfection.

[0053] 3. Washing: Add 10-50 microliters of washing liquid to each active well, wash 5-8 times, shake for 5 seconds each time, suck out the washing liquid (discard after disinfection), and pat dr...

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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 present invention relates to the technical fields of molecular biology, biomedicine, and clinical immunoassay, 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 a 1997 report of the World Health Organization, the number of deaths from cancer patients worldwide is as high as 6.6 million annually, and by 2020 the number of cancer patients worldwide will reach 15 million. Various malignant tumors seriously threaten human health. In the early stages of tumor onset, there are often no symptoms, and once the symptoms are discovered, it has reached the middle and late stages that cannot be treated. Today, when the pathogenesis and treatment of tumors have not been completely resolved, to reduce the mortality of tumor patients, it is necessary to carry out general tumor surveys at the basic level to achieve early detection and early...

Claims

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Application Information

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Patent Type & Authority Applications(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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