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A biochip that can expand the scope of immunodetection

A technology of immunoassay and biochip, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low antigen concentration inaccuracy, complicated operation, etc., achieve improved detection range and precision, high sensitivity, and easy operation Effect

Active Publication Date: 2020-08-04
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

However, the disadvantage of this method is that it is necessary to determine whether the antigen concentration in the sample is too high through a one-step test before using this method
Using this method without prior judgment can lead to inaccurate detection of low antigen concentrations
Pre-judgment will add a repeated measurement, and the operation is complicated

Method used

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  • A biochip that can expand the scope of immunodetection
  • A biochip that can expand the scope of immunodetection
  • A biochip that can expand the scope of immunodetection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Preparation of magnetic sensitive NT-proBNP biochip

[0051] Experimental Materials

[0052] NT-proBNP capture antibody I (affinity rate is about 5×10 9 M -1 )

[0053] NT-proBNP capture antibody II (affinity rate is about 5×10 8 M -1 )

[0054] Magnetic particle labeled antibody NT-proBNP antibody III (can be paired with capture antibody I and capture antibody II at the same time)

[0055] Conventional reagents are commercially available

[0056] (1) Biochip table functionalization

[0057] Treat the surface of single crystal silicon chip with oxygen plasma for 10 minutes, then immerse it in anhydrous ethanol solution containing 2-10% ethoxysilane and place it in a 37℃ shaker for 0.5-2h, and then use anhydrous Washed with ethanol for 5 times, and finally dried with nitrogen. At this time, the surface of the chip has been aminated, and an amino biochip is prepared.

[0058] (2) Connection of fixed carrier

[0059] Soak the amino silicon wafer in phosphate buffer containin...

Embodiment 2

[0062] Example 2: Determination of NT-proBNP detection range

[0063] Prepare NT-proBNP antigen into 0pg / mL, 15pg / mL, 50pg / mL, 100pg / mL, 500pg / mL, 1000pg / mL, 5000pg / mL, 10000pg / mL and 40000ng / mL antigen standard series, and measure them respectively Different standard products, get the corresponding magnetic signal intensity (ie the change in magnetic resistivity), and fit the standard curves A and B of antibody I and antibody II respectively according to the antigen concentration and magnetic signal intensity, such as Figure 4 Shown in A and 4B. The formulas of NT-proBNP antigen concentration-magnetoresistance change rate are: standard curve A, y=0.263 (x^0.595); standard curve B, y=0.103 (x^0.590). And draw the two standard curves together, such as Figure 4 Shown in C.

[0064] Antibody I alone detects NT-proBNP, the detection range is 15pg / ml to 5000pg / ml (magnetoresistance signal range is 0.14-4.10). Antibody II alone detects NT-proBNP, the detection range is 100pg / ml to 40...

Embodiment 3

[0066] Example 3: Determination of actual NT-proBNP samples

[0067] The 53 actual human NT-proBNP plasma samples whose concentrations have been determined by Roche Cobas e411 were used as the test samples, and the magnetic-sensitive NT-proBNP biochip was used for detection, and the detection concentrations were calculated using the two standard curves in Example 4. Select 34 samples in the concentration range of 100pg / ml to 5000pg / ml in the area where the hyperbola intersects for analysis. Compare the consistency of the magnetic sensitive immunochip Roche Cobas e411 in detecting the concentration of NT-proBNP in human plasma, such as Figure 5 Shown.

[0068] The results of the methodological comparison show that using a single standard A and B, the correlation coefficient r is 0.980 and 0.982, respectively. However, using the double standard curve method, the correlation coefficient r is 0.991. The precision of NT-proBNP detection has been greatly improved. The detection perfo...

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Abstract

The invention discloses a biochip being able to widen the immunodetection range. The biochip comprises two or above distinguishable detection areas, the detection areas can immobilize an antibody I and an antibody II, and the antibody I and the antibody II are combined with a same antigen. The chip has the advantages of simplicity in operation, high sensitivity, wide detection range, effective widening of the immunodetection range of a sample, and improvement of the detection precision.

Description

Technical field [0001] The invention relates to the technical field of in vitro immunoassays, in particular to a biochip that can expand the detection range. Background technique [0002] Solid-phase immunoassay is a technique commonly used in immunoassays. It is a detection technique based on immune response, which is highly efficient and sensitive. In the solid-phase immunoassay, the solid-phase material is used as a carrier of immunoreagents and as a place for immune response. Biochip is a detection method based on solid-phase immunotechnology. The immune reaction occurs on the solid-phase surface, which is often limited by surface steric hindrance and the nature of the antigen and antibody itself, resulting in a narrow detection range and low detection precision, which is difficult to meet the actual requirements. Due to the needs of applications, it is necessary to broaden the detection range of solid phase immunoassay to meet the needs of sample detection. [0003] Accordin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543
Inventor 高云华霍卫松石西增
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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