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IGF family protein combined detection chip

A combined detection and protein technology, applied in the field of biomedicine, can solve the problems of cumbersome operation, single detection index, and low sample consumption.

Active Publication Date: 2014-04-02
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the deficiencies of the prior art, such as cumbersome operation, single detection index and low sensitivity, the purpose of the present invention is to provide a new protein joint detection chip for the insulin-like growth factor (IGF) signaling pathway, so as to realize the high detection of IGF family proteins. Throughput, high sensitivity, high specificity and low cost detection, with the advantages of cheap, convenient, sensitive, accurate, high throughput, less specimen consumption, and can be popularized and scaled in ordinary laboratories

Method used

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Examples

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Embodiment 1

[0025] Example 1: Construction of the IGF family protein combined detection chip of the present invention

[0026] In order to develop the IGF family protein combined detection chip of the present invention, firstly select existing antibody products from suppliers or develop appropriate antibody pairs by themselves. These antibody pairs are then used to create a chip to simultaneously detect 10 proteins of the IGF family.

[0027] material

[0028] All antibodies were purchased from R&D Systems (Minneapolis, Minnesota, USA) or prepared by RayBiotech (Norcross, Georgia, USA). IGF family proteins come from R&D company or prepared by RayBiotech company. Streptavidin was purchased from BD PharMingen. The slides were purchased from Corning (Corning, New York, USA).

[0029] Table 1: Sources of specific antibodies

[0030]

[0031] method

[0032] A non-contact Piezorray Arrayer spotting machine from PerkinElmer (Shelton, Connecticut, USA) was used to spot the capture antibody on the glass...

Embodiment 2

[0050] Example 2: The IGF family protein combined detection chip of the present invention is used for the detection of liver cancer tissue samples and their adjacent tissue samples

[0051] In order to verify the application potential of the IGF family protein combined detection chip of the present invention, the cancer tissues and adjacent tissues of 25 patients were tested in this example, and the expression of 10 proteins from the IGF signaling pathway in these tissues was analyzed. Level.

[0052] Sample preparation

[0053] The cancer tissue samples and their adjacent tissue samples (3-5 cm from the cancer tissue) were collected from 25 patients who were diagnosed as having hepatocellular carcinoma by surgery at the First Affiliated Hospital of Sun Yat-sen University. All procedures are in compliance with the health guidelines of the national institution, and approved by the institutional review board and the hospital ethics committee and the patient's consent. The diagnosis, ...

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Abstract

The invention relates to an insulin-like growth factor (IGF) family protein combined detection chip. The chip includes: glass slides coated by active amino propyl silane; multiple micro-arrays fixed on each glass slide by a chip dot matrix; and gaskets for separating the micro-arrays into different hybrid areas, wherein each of the micro-arrays contains a specific antibody corresponding to an IGF family protein. The chip also includes the following reaction reagents: an biotin-marked detection antibody mixed solution, a closed solution which adopts a phosphate buffer solution containing 5% bovine serum albumin, a standard protein mixture, and streptavidin marked by adopting a fluorescent dye HyLight555 with the excitation wavelength of 550 nm and the emission wavelength of 566 nm. The IGF family protein combined detection chip can complete multi-detection indicator, high-throughput and high-sensitivity detection which cannot be completed by traditional methods such as ELISA, western blot and the like, and is quite suitable for multi-link simultaneous detection of mass target proteins of an IGF-1 signal pathway.

Description

Technical field [0001] The invention relates to the field of biomedical technology, and relates to a protein combined detection chip, in particular to a combined detection chip for insulin-like growth factor (IGF) family proteins. Background technique [0002] Insulin-like Growth Factor (IGF) signaling system plays an important physiological role in regulating cell proliferation, differentiation and apoptosis through the interaction with specific receptors on cell membrane. The IGF signaling system consists of the following three parts: ligand (IGF-1, IGF-2 and insulin), receptor (IGF-1R, IGF-2R, insulin receptor (IR), IGF-1R / IR hybrid receptor (HR) etc.) and six IGF high-affinity binding proteins (IGFBP1-6). Among them, IGF-1 and IGF-2 play an important role in fetal development and postnatal life through endocrine, paracrine and autocrine mechanisms. The mitotic, differentiation and anti-apoptotic properties of IGF are mainly mediated by IGF-1R. When IGF-1 or IGF-2 binds to ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6863G01N2333/4745
Inventor 黄若磐毛应清陈蕴如
Owner RAYBIOTECH INC GUANGZHOU
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