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Process for preparing antigen microarray based on self antibody repertoire

A technology of autoantibodies and production methods, which is applied in the field of biotechnology microarray production, can solve problems such as poor method sensitivity, method repeatability interference, isotope pollution, etc., and achieve strong practical and industrial development value, strong practicability, Reasonable effect of technical design

Inactive Publication Date: 2002-04-10
SOUTHEAST UNIV
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Problems solved by technology

At present, the detection methods for autoantibodies include colloidal gold immunofiltration technology, enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA). The former has poor sensitivity, while the latter has isotope contamination and instability. and other characteristics, it is always difficult to be satisfactory in use, and the ELISA technology widely used in clinical laboratories at present, its method repeatability is easily interfered by various measurement factors, and the above method is only applicable to the determination of a single item (i.e. a This experiment can only detect one autoantibody at a time), so it is necessary to find a sensitive, stable and high-throughput detection method

Method used

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Embodiment Construction

[0017] ① Pretreatment of the substrate (glass slide): meet the requirements suitable for molecular assembly (including cleaning requirements, roughness requirements, etc.).

[0018] ② Put the clean hydroxylated surface-treated glass slides in a certain concentration of APTES (2%) acetone solution. After soaking for an adaptation time, take them out and rinse them with acetone and ether in turn, and dry them naturally for later use.

[0019] ③ Make a gel molecular layer (agarose) on the surface of the substrate → NalO 4 Oxidation for 20 minutes→washing with distilled water for 30 minutes→drying and then use a certain concentration of GA to make the aldylated surface, and dry it in an oven for later use.

[0020] ④ Use spotting equipment to spot different antigen molecules (conditions pre-explored) on the glass slide, the spotting concentration is 100 μg / ml, and the spotting volume is 0.1 μl. Anti-human gamma globulin and bovine serum albumin (BSA) are set as the Positive and n...

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Abstract

A process for preparing an antigen microarray based on self antibody repertoire includes pre-treating substrate for adapting molecular self-assembly, generating a layer of agarose gel moleculae, chemical modifying it, designing antigen microarray, preparing antigen microarray by applying different biomacromolecular antigens on the surface of said substrate, and testint the chip by use of microscope or fluorescent scan to acquire the information about specimen. Its test method includes culturing the said microarray chip and specimen to be tested in warmhouse, hybridization with the second antigen probe of three fluorescein labels, and acquiring the information about specimen by multi-channel fluorescent scan.

Description

1. Technical field [0001] The invention relates to a method for making a biotechnology microarray, in particular to a method for making a microarray based on an autoantibody spectrum antigen. 2. Background technology [0002] Biochip is a combination of planar microfabrication and biomolecular self-assembly technology, and assembles a large number of different biomolecular microarrays on a tiny glass slide to achieve large-scale detection of target DNA or protein molecular information. This technology is considered to be an efficient tool in life science research, so it has become a hot spot in the development of science and technology at home and abroad. [0003] The protein immunoassay chip is different from the general DNA chip detection technology. Its principle is mainly designed based on the specific binding reaction between antigen and antibody. The target molecule tested has only structural specificity, but no sequence specificity. . At present, the detection metho...

Claims

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Application Information

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IPC IPC(8): C07K17/14G01N33/50G01N33/53G01N33/68
CPCB82Y15/00B82Y30/00
Inventor 虞伟顾宁武建国廖建辉张海黔
Owner SOUTHEAST UNIV
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