Antibody and magnetic bead directed coupling method

A magnetic bead and antibody technology, which is applied in the field of biotechnology detection, can solve the problems of reducing the sensitivity and specificity of the reagent, not exceeding 25%, and low labeling efficiency, and achieves the improvement of the sensitivity of the reagent, the increase of the yield, and the thin hydration layer. Effect

Pending Publication Date: 2019-12-13
山东康华生物医疗科技股份有限公司
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, by using different types of reducing agents, different reducing times and different reducing conditions to screen the antibody reduction scheme, the yield of the final target product does not exceed 25%, and the labeling efficiency is extremely low.
This method easily causes the interference of rheumatoid factor (RF) in addition, reduces the sensitivity and specificity of reagent

Method used

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  • Antibody and magnetic bead directed coupling method
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Examples

Experimental program
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Effect test

Embodiment 1

[0025] A test kit for detecting VEGF prepared by the directional coupling method of the present invention comprises the following components: magnetic microsphere suspension coated with VEGF antibody (the working concentration of the magnetic bead microsphere is 0.1mg / ml- 0.35mg / ml, the concentration of VEGF antibody coated on the surface of magnetic beads is 10-50ug / mg), VEGF reference product (VEGF reference product is prepared from VEGF standard product and reference product dilution, the concentrations of VEGF reference product are respectively 0pg / mL, 30pg / mL, 125pg / mL, 500pg / mL, 1000pg / mL, 2000pg / mL;), VEGF quality control substance (VEGF reference substance is prepared from VEGF standard substance and reference substance dilution, VEGF quality control substance The concentration of control product is 125pg / mL, 1000pg / mL), VEGF enzyme conjugate (VEGF enzyme conjugate is the VEGF monoclonal antibody of horseradish peroxidase labeling, and the concentration of VEGF enzyme c...

Embodiment 2

[0035] A test kit for detecting VEGF prepared by the directional coupling method of the present invention comprises the following components: magnetic microsphere suspension coated with VEGF antibody (the working concentration of the magnetic bead microsphere is 0.1mg / ml- 0.35mg / ml, the concentration of VEGF antibody coated on the surface of magnetic beads is 10-50ug / mg), VEGF reference product (VEGF reference product is prepared from VEGF standard product and reference product dilution, the concentrations of VEGF reference product are respectively 0pg / mL, 30pg / mL, 125pg / mL, 500pg / mL, 1000pg / mL, 2000pg / mL;), VEGF quality control substance (VEGF reference substance is prepared from VEGF standard substance and reference substance dilution, VEGF quality control substance The concentration of control product is 125pg / mL, 1000pg / mL), VEGF enzyme conjugate (VEGF enzyme conjugate is the VEGF monoclonal antibody of horseradish peroxidase labeling, and the concentration of VEGF enzyme c...

Embodiment 3

[0045] A test kit for detecting VEGF prepared by the directional coupling method of the present invention comprises the following components: magnetic microsphere suspension coated with VEGF antibody (the working concentration of the magnetic bead microsphere is 0.1mg / ml- 0.35mg / ml, the concentration of VEGF antibody coated on the surface of magnetic beads is 10-50ug / mg), VEGF reference product (VEGF reference product is prepared from VEGF standard product and reference product dilution, the concentrations of VEGF reference product are respectively 0pg / mL, 30pg / mL, 125pg / mL, 500pg / mL, 1000pg / mL, 2000pg / mL;), VEGF quality control substance (VEGF reference substance is prepared from VEGF standard substance and reference substance dilution, VEGF quality control substance The concentration of control product is 125pg / mL, 1000pg / mL), VEGF enzyme conjugate (VEGF enzyme conjugate is the VEGF monoclonal antibody of horseradish peroxidase labeling, and the concentration of VEGF enzyme c...

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Abstract

The invention belongs to the field of biotechnology detection and relates to an antibody and magnetic bead directed coupling method. The method includes the following steps of: (1) magnetic bead pretreatment; (2) S-NHS treatment; (3) EDC treatment; and (4) antibody addition. With the method provided by the invention adopted, the directed coupling of antibodies and magnetic beads can be realized; the yield of effective antibodies is greatly improved; and the actual sensitivity and accuracy of detection are improved.

Description

technical field [0001] The invention relates to the field of biotechnology detection, in particular to a method for directional coupling of antibodies and magnetic beads. Background technique [0002] Chemiluminescent immunodiagnostic technology uses chemiluminescent agents, catalytic luminescent enzymes and other substances to label antigens or antibodies. Redox reactions. The visible light released in the reaction is finally collected and processed by supporting equipment to obtain the concentration of the measured substance, so as to achieve the purpose of measurement. Compared with traditional immunological techniques, chemiluminescence has the advantages of high degree of automation, good specificity, high precision, and wide detection range. [0003] During the development and preparation of magnetic particle chemiluminescent reagents, there are many coupling methods for antibodies and magnetic beads, but there is a problem in all of them, that is, the direction in w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/535G01N33/543
CPCG01N33/53G01N33/535G01N33/54326
Inventor 杨致亭田永帅金宇婷隋振国杨锋斌葛玮
Owner 山东康华生物医疗科技股份有限公司
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