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Biochip utilizing hook effect to enlarge detection range and detection method thereof

A hook-like effect and biochip technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as inflexibility, and achieve the effects of high detection sensitivity, short detection time, and easy operation

Active Publication Date: 2016-08-03
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method saves time, but it is not flexible enough due to the physical location of the detection area and indication area

Method used

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  • Biochip utilizing hook effect to enlarge detection range and detection method thereof
  • Biochip utilizing hook effect to enlarge detection range and detection method thereof
  • Biochip utilizing hook effect to enlarge detection range and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1: Structure of biochip and test card comprising biochip

[0052] refer to image 3 , the biochip 310 of the present invention includes a target region 320 immobilized with a capture antibody, and an indicator region 330 immobilized with an antigen. The substrate 300 of the biochip has a sample loading hole 340, a microchannel 350, a liquid waste area 360 and an exhaust hole 370, and the target area 320 and the indicator area 330 are located side by side in the microchannel 350 (see image 3 ).

[0053] The width of the microporous channel is 10-200 microns, the height is 10-200 microns, and the length is 3-5 cm.

[0054] The material for making the microchannel can be silicon wafer, metal, glass, ceramics, organic polymers such as plastic, rubber, etc. Preferably, the material for making the microchannel is plastic.

[0055] The manufacturing process of the microchannel can be lithography, chemical etching technology, photolithography electroforming injec...

Embodiment 2

[0058] Embodiment 2: the preparation of magnetically sensitive C-reactive protein (CRP) biochip

[0059] Experimental Materials

[0060] CRP capture antibody, CRP antigen molecule, magnetic particle labeled CRP antibody;

[0061] Conventional reagents are commercially available

[0062] (1) Functionalization of biochip surface

[0063] Use oxygen plasma to treat the surface of a single crystal silicon chip for 10 minutes, then immerse it in an absolute ethanol solution containing 2-10% ethoxysilane and place it in a shaker at 37 degrees Celsius for 0.5-2 hours, and then use Wash with absolute ethanol for 5 times, and finally blow dry with nitrogen. At this time, the surface of the chip has been aminated, that is, the amino biochip is prepared.

[0064] (2) Connection of fixed carrier

[0065] Amino silicon chips were soaked in phosphate buffer containing 5% glutaraldehyde, reacted at room temperature for 1 hour, and then washed with phosphate buffer, then CRP capture antib...

Embodiment 3

[0072] Example 3: Determination of CRP detection hook effect and determination of detection range

[0073] CRP antigen formulated at 0, 0.003, 0.006, 0.012, 0.025, 0.049, 0.098, 0.195, 0.391, 0.552, 0.781, 1.105, 1.563, 3.125, 6.25, 12.5, 25, 50, 100, 200, 350, 450 μg / mL Antigen standard series, measure the standard products with different concentrations, obtain the corresponding magnetic signal intensity (ie, the change value of magnetoresistivity), and fit the "target area" and "indicator area" according to the antigen concentration and magnetic signal intensity. "Standard curves A and B, the standard curve adopts classic four-parameter fitting (LogisticEquation), and the equation is: y=A 2 +(A 1 -A 2 ) / (1+(x / x 0 )^p), such as Figure 4 As shown in A and 4B, fitting the two figures together yields Figure 4 c.

[0074] Using sandwich immunoassay to detect CRP, the detection range is 0.003μg / mL-0.781μg / mL, when the antigen concentration reaches 0.781μg / mL, the "target a...

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Abstract

The invention discloses a biochip utilizing the hook effect to enlarge the detection range and a detection method thereof. The biochip comprises at least one target area, a capturing antibody is fixed in the target area, the capturing antibody is combined with an analyte, through the labeling antibody combined with the analyte, detectable signals can be generated and can be used to quantitatively analyze the analyte in a sample; the biochip also comprises at least one indicating area, an antigen is fixed in the indicating area; and the fixed antigen is not combined with the analyte and is combined with labeling antibody that is not combined with the analyte to generate a detectable signal, which can be used to judge whether a hook effect is generated or not. The biochip has the advantages of flexible and convenient operation, time saving, and high sensitivity, largely enlarges the detection range of biological molecules, and avoids the false negative results.

Description

technical field [0001] The invention relates to the technical field of in vitro immune analysis, in particular to a biochip and a detection method thereof, which can significantly expand the detection range of biomolecules. Background technique [0002] The antigen-antibody reaction refers to the specific binding reaction between the antigen and the corresponding antibody. It can occur both inside the body and outside the body. The antigen-antibody reaction in the body is the effect of the humoral immune response, and the combination of antigen and antibody in vitro is mainly used for the detection of antigen or antibody and for immunological diagnosis. Immunoassay is an analytical method that uses antigen-antibody specific binding reactions to detect the presence or concentration of drugs, hormones, proteins, microorganisms and other substances in complex samples. This analysis is based on the property of an antibody that it specifically binds only one or a very limited n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 高云华张磊石西增
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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