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SDF-1 determination kit and preparation and detection method thereof

A technology of SDF-1 and reagent kit, which is applied in the field of immunological determination, can solve the problems of many influencing factors, high price, and long measurement time, and achieve the effect of improving detection precision, high degree of automation, and small amount of detection samples

Pending Publication Date: 2022-03-11
ANHUI DAQIAN BIO ENG LIMITED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the enzyme-linked immunosorbent assay has high detection sensitivity, but its operation process is complicated, the measurement time is long, and there are many influencing factors, and this method is mainly used in scientific research and cannot be used in clinical diagnosis.
However, the currently commercially available SDF-1 detection ELISA kits are mainly imported from abroad, and the price is expensive, which has seriously affected the wide application of this product.
Immunohistochemistry has the advantages of good specificity and high sensitivity, but there are many uncontrollable factors in the experimental process, such as the choice of primary antibody, the incubation time of secondary antibody, and the difference in color development time, etc., which will cause inaccurate results. stable, and may even lead to different or opposite results in different labs
Western blotting has the advantages of good specificity, but its disadvantages are poor repeatability and antibody titer needs to be titrated, etc., and monoclonal antibodies cannot recognize denatured antigens
[0004] However, most of the SDF-1 currently on the global market is produced by prokaryotic cell fusion expression technology, or the traditional process of denaturation and renaturation, the product purity is low, and the biological activity is difficult to meet the needs of basic research; -1 During activity detection, a small amount of small antigen-antibody complexes are difficult to form turbidity, which does not meet the requirements for trace quantification

Method used

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  • SDF-1 determination kit and preparation and detection method thereof
  • SDF-1 determination kit and preparation and detection method thereof
  • SDF-1 determination kit and preparation and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] A SDF-1 assay kit in this embodiment includes reagent R1, reagent R2 and calibrator.

[0052] Reagent R1 includes: phosphate buffer 10mmol / L (pH7.4), potassium sorbate 0.05g / L, PEG-400030g / L, sodium chloride 5g / L, Tween-40 2mL / L.

[0053] Reagent R2 includes: phosphate buffer 40mmol / L (pH7.4), potassium sorbate 1g / L, sodium chloride 0.5g / L, Tween-40 2mL / L, anti-human SDF-1 antibody mixed latex particles 30mg / L; Wherein, the latex particles in the anti-human SDF-1 antibody mixed latex particles use latex particles of two different particle sizes (100nm, 180nm), and the anti-human SDF-1 antibody uses a mouse anti-human SDF-1 monoclonal antibody.

[0054] Calibrators include: MOPS buffer 100mmol / L (pH7.3), BSA 3g / L, sodium azide 0.5g / L, 200~3200U / mL SDF-1.

[0055] The preparation method of the kit of this embodiment:

[0056] (1) Preparation of reagent R1:

[0057] According to the component content of the reagent R1, the components are mixed in the same container, an...

Embodiment 2

[0075] A SDF-1 assay kit in this embodiment includes reagent R1, reagent R2 and calibrator.

[0076] Reagent R1 includes: glycine buffer 100mmol / L (pH7.8), sodium nitrite 1g / L, dextran sulfate sodium 40g / L, EDTA 15g / L, Span80 5mL / L.

[0077] Reagent R2 includes: glycine buffer 100mmol / L (pH7.8), sodium nitrite 2g / L, EDTA1g / L, Span805mL / L, anti-human SDF-1 antibody mixed latex particles 50mg / L; among them, anti-human SDF- 1 The latex particles in the antibody mixed latex particles use two types of latex particles with different particle sizes (180nm, 200nm), and the anti-human SDF-1 antibody uses a mouse anti-human SDF-1 monoclonal antibody.

[0078] Calibrators include: MOPS buffer 100mmol / L (pH7.3), BSA 3g / L, sodium azide 0.5g / L, 200~3200U / mL SDF-1.

[0079] The preparation method of the kit of this embodiment:

[0080] (1) Preparation of reagent R1:

[0081] According to the component content of the reagent R1, the components are mixed in the same container, and after mix...

Embodiment 3

[0099] A SDF-1 assay kit in this embodiment includes reagent R1, reagent R2 and calibrator.

[0100] Reagent R1 includes: Tris-HCl buffer solution 50mmol / L (pH 7.5), sodium azide 0.5g / L, PEG-600050g / L, BSA 10g / L, Triton-100 3.5mL / L.

[0101]Reagent R2 includes: MOPS buffer 50mmol / L (pH 7.5), procline-300 1.0g / L, BSA 0.5g / L, Triton-100 3.5mL / L, anti-human SDF-1 antibody mixed latex particles 40mg / L L; Among them, the latex particles in the anti-human SDF-1 antibody mixed latex particles use latex particles with two different particle sizes (150nm, 190nm), and the anti-human SDF-1 antibody uses a mouse anti-human SDF-1 monoclonal antibody.

[0102] Calibrators include: MOPS buffer 100mmol / L (pH7.3), BSA 3g / L, sodium azide 0.5g / L, 200~3200U / mL SDF-1.

[0103] The preparation method of the kit of this embodiment:

[0104] (1) Preparation of reagent R1:

[0105] According to the component content of the reagent R1, the components are mixed in the same container, and after mixing...

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Abstract

The invention provides an SDF-1 determination kit. The SDF-1 determination kit comprises a reagent R1 and a reagent R2, the reagent R1 comprises 10-100 mmol / L of a buffer solution, 0.05-1 g / L of a preservative, 30-50 g / L of a coagulant, 5-15 g / L of a stabilizer and 2-5 mL / L of a surfactant; the reagent R2 comprises 40-100 mmol / L of a buffer solution, 1-2 g / L of a preservative, 0.5-1 g / L of a stabilizer, 2-5 mL / L of a surfactant and 30-50 mg / L of anti-human SDF-1 antibody mixed latex particles; wherein latex particles in the anti-human SDF-1 antibody mixed latex particles adopt two different particle sizes. The invention also provides a preparation method and a determination method of the SDF-1 determination kit. The kit is used for measuring the content of SDF-1, is simple and convenient to operate, short in detection time and low in cost, and also has relatively high sensitivity, precision and stability.

Description

technical field [0001] The invention relates to the field of immunological determination, in particular to an SDF-1 determination kit and a preparation and detection method thereof. Background technique [0002] Stromal cell derived factor-1 (SDF-1) is a newly discovered small molecule cytokine, which belongs to the CXC (Cys-X-Cys) class of chemokine proteins and is named CXCL12 systematically. It is the only known natural chemokine that can bind to and activate the receptor CXCR4, and it has two forms: "SDF-1α / CXCL12a and SDF-1β / CXCL12b". SDF-1 and CXCR4 are widely expressed in a variety of cells and tissues, including immune cells, brain, heart, kidney, liver, lung and spleen, which play a critical role in the development of the immune system, circulatory system and central nervous system important role. At the same time, SDF-1 is closely related to HIV (Human immunodeficiency virus) infection, inflammatory response, regulation of hematopoietic cells, and migration of tu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6863G01N33/54313G01N2333/521
Inventor 芮双印梁艳
Owner ANHUI DAQIAN BIO ENG LIMITED
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