Fluorescent microsphere labeled antibody as well as preparation method and application thereof

A technology for labeling antibodies and fluorescent microspheres, which is applied in the field of immunoassays, can solve the problems of different cost-effectiveness of antibodies or antigens, fixed production or inventory, long transportation cycle, etc., and achieve good sensitivity and specificity, sensitivity and specificity Retention, Sensitivity, and Specificity Consistent Effects

Pending Publication Date: 2020-08-25
北京康思润业生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specific antibodies or antigens that are currently used in immune labeling, due to the different production processes and levels of manufacturers, the cost performance of antibodies or antigens are different. Although high-quality antibodies or antigens have high specificity and sensitivity, they are also expensive. Relatively expensive, t...

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  • Fluorescent microsphere labeled antibody as well as preparation method and application thereof
  • Fluorescent microsphere labeled antibody as well as preparation method and application thereof
  • Fluorescent microsphere labeled antibody as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1.1 Preparation of fluorescent microsphere-labeled antibody

[0033] 1.1.1 Microsphere cleaning:

[0034]The original concentration of the fluorescent microspheres was 2.0wt%, and they were centrifuged and washed with 10mM phosphate buffer (pH6.0) at a rotation speed of 14000rpm for 10min. The supernatant was discarded and repeated twice to obtain microsphere precipitates.

[0035] 1.1.2 Activation:

[0036] 1) Preparation of chemical crosslinking agent solution:

[0037] The chemical cross-linking agent is N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide with a weight ratio of 1:1, respectively using 10mM (pH6.0) Phosphate buffer solution prepared into a 500mg / ml solution for subsequent use;

[0038] 2) Activation of fluorescent microspheres:

[0039] Add 10mM phosphate buffer solution (pH6.0) to the washed microsphere precipitate, and ultrasonically mix (40kHz, 6min) to obtain a 100mg / mL fluorescent microsphere solution. The 500mg / ml N Mixe...

Embodiment 2

[0072] 2.1 Preparation of fluorescent microsphere-labeled antibody

[0073] 2.1.1 Microsphere cleaning:

[0074] The original concentration of the fluorescent microspheres was 2.0 wt%, and they were centrifuged and washed with 10 mM phosphate buffer (pH6.0) at a rotation speed of 14000 rpm for 10 min. The supernatant was discarded and repeated twice to obtain microsphere precipitates.

[0075] 2.1.2 Activation:

[0076] 1) Preparation of chemical crosslinking agent solution:

[0077] The chemical cross-linking agent is N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide with a weight ratio of 1:1, respectively using 10mM (pH6.0) Phosphate buffer solution prepared into a 500mg / ml solution for subsequent use;

[0078] 2) Activation of fluorescent microspheres:

[0079] Add 10mM phosphate buffer solution (pH6.0) to the washed microsphere precipitate, and ultrasonically mix (40kHz, 6min) to obtain a 10mg / mL fluorescent microsphere solution. The 500mg / ml N Mi...

Embodiment 3

[0116] 3.1 Preparation of fluorescent microsphere-labeled antibody

[0117] 3.1.1 Microsphere cleaning:

[0118] The original concentration of the fluorescent microspheres was 2.0 wt%, and they were centrifuged and washed with 10 mM phosphate buffer (pH6.0) at a rotation speed of 14000 rpm for 9 minutes. The supernatant was discarded and repeated twice to obtain microsphere precipitates.

[0119] 3.1.2 Activation:

[0120] 1) Preparation of chemical crosslinking agent solution:

[0121]The chemical cross-linking agent is N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide with a weight ratio of 1:1, respectively using 10mM (pH6.0) Phosphate buffer solution prepared into a 500mg / ml solution for subsequent use;

[0122] 2) Activation of fluorescent microspheres:

[0123] Add 10mM phosphate buffer solution (pH6.0) to the washed microsphere precipitate, and ultrasonically mix (40kHz, 6min) to obtain a 100mg / mL fluorescent microsphere solution. The 500mg / ml ...

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Abstract

The invention relates to a fluorescent microsphere labeled antibody as well as a preparation method and application thereof. The preparation method comprises the following steps: adding 100-1000mg/mLof a chemical cross-linking agent solution into 1-50mg/mL of a fluorescent microsphere solution for activation; mixing the labeled antibody with quality control protein to obtain total protein, and adding the total protein into the obtained activated product for reaction; and adding a sealing agent into the obtained solution for sealing, and centrifuging to obtain the fluorescent microsphere labeled antibody. The labeled antibody/antigen and other proteins are simultaneously labeled on the surface of the marker, thereby lowering the consumption of the labeled antibody/antigen and lowering thecost; and moreover, the original performance of the labeled antibody/antigen can be kept good and stable.

Description

technical field [0001] The invention relates to a fluorescent microsphere-labeled antibody and its preparation method and application, belonging to the technical field of immune detection. Background technique [0002] When detecting certain antigens or specific proteins, immunolabeling techniques are usually applied. Immunolabeling techniques (immunolabelling techniques) refer to the use of fluorescein, radioactive isotopes, enzymes, ferritin, colloidal gold and chemical (or bio) luminescent agents as tracers to label antibodies or antigens for antigen-antibody reactions. , Enzyme label detector and other instruments, direct microscopic observation or automatic determination of the experimental results, can conduct qualitative and positioning research on antigen-antibody reactions at the cellular, subcellular and molecular levels; or apply various liquid and solid phases Immunoassay method for qualitative and quantitative determination of haptens, antigens or antibodies in...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/58G01N33/558
CPCG01N33/533G01N33/558G01N33/582G01N33/587
Inventor 张军李海燕
Owner 北京康思润业生物技术有限公司
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