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Soluble leptin receptor (sOB-R) latex reinforced immunonephelometry assay kit by utilizing surface functional group

A technology for detecting kits and surface functional groups, which can be used in biological testing, material inspection, etc. It can solve the problems of poor control of sample dilution, narrow linear range, and complicated operation, and achieve shortened reaction time, increased sensitivity, and extended validity period. Effect

Active Publication Date: 2015-03-25
CHONGQING ZHONGYUAN BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the sensitivity of ELISA is high, its operation is complicated and takes more time. Generally, the results are qualitative or semi-quantitative. The deviation of quantification is large, and the linear range is narrow. It is not easy to control the dilution of samples.

Method used

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  • Soluble leptin receptor (sOB-R) latex reinforced immunonephelometry assay kit by utilizing surface functional group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Preparation of R1 reagent:

[0042] Weigh 1.9g stachyose, 0.5g alum, 1.9g sodium fructose diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycine, 20g NaCl, 50g PEG8000, 0.5g sodium azide, 25g bovine serum albumin and 14.2g Dissolve EDTA in 0.8L of distilled water, adjust the pH to 7.4, and set the volume to 1L to obtain reagent R1.

[0043] Preparation of R2 reagent:

[0044] Add PBS buffer solution with a pH of 7.4 and a concentration of 100mmol / L to the mouse anti-human sOB-R antibody for dialysis at 4°C, and complete the dialysis after changing the water 3 times in the middle. -R antibody diluted to a concentration of 2mg / ml mouse anti-human sOB-R antibody diluent; the polystyrene latex microspheres whose surface functional groups are carboxyl groups were washed with distilled water.

[0045] The washed polystyrene latex microspheres were then diluted to a mass concentration of 1% with PBS buffer solution having a pH of 7.4 and a concentration of 100 mmol / L. Add...

Embodiment 2

[0053] Preparation of R1 reagent:

[0054] Weigh 1.9g stachyose, 0.5g alum, 1.9g sodium fructose diphosphate, 0.3g sodium hexametaphosphate, 1.8g glycine, 9g NaCl, 15g PEG8000, 0.5g sodium azide, 5g bovine serum albumin and 14.5g Dissolve EDTA in 0.8L of distilled water, adjust the pH to 7.4, and set the volume to 1L to obtain reagent R1.

[0055] Preparation of R2 reagent:

[0056] Add PBS buffer solution with a pH of 7.4 and a concentration of 100mmol / L to the mouse anti-human sOB-R antibody for dialysis at 4°C, and complete the dialysis after changing the water 3 times in the middle. -R antibody diluted to a concentration of 2mg / ml mouse anti-human sOB-R antibody diluent; the polystyrene latex microspheres whose surface functional groups are carboxyl groups were washed with distilled water.

[0057] The washed polystyrene latex microspheres were then diluted to a mass concentration of 1% with PBS buffer solution having a pH of 7.4 and a concentration of 100 mmol / L. Add 0...

Embodiment 3

[0061] Preparation of R1 reagent:

[0062] Weigh 1.8g glycine, 9g NaCl, 15g PEG8000, 0.5g sodium azide, 5g bovine serum albumin and 14.5g EDTA, dissolve in 0.8L distilled water, adjust the pH to 7.4, and dilute to 1L to obtain reagent R1.

[0063] Preparation of R2 reagent:

[0064] Add PBS buffer solution with a pH of 7.4 and a concentration of 100mmol / L to the mouse anti-human sOB-R antibody for dialysis at 4°C, and complete the dialysis after changing the water 3 times in the middle. -R antibody diluted to a concentration of 2mg / ml mouse anti-human sOB-R antibody diluent; the polystyrene latex microspheres whose surface functional groups are carboxyl groups were washed with distilled water.

[0065] The washed polystyrene latex microspheres were then diluted to a mass concentration of 1% with PBS buffer solution having a pH of 7.4 and a concentration of 100 mmol / L. Add 0.01g of EDC to 1L of the above latex dilution, stir and react at room temperature for 30min, centrifuge...

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Abstract

The invention relates to the field of in-vitro diagnostic reagents, and particularly relates to a soluble leptin receptor (sOB-R) latex reinforced immunonephelometry assay kit by utilizing a surface functional group. The assay kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is prepared from a buffer solution 1, a stabilizing agent 1, a preservative 1, EDTA, a gelling agent and a protective agent 1; the reagent R2 is prepared from polystyrene latex microspheres crosslinked with an sOB-R antibody, a buffer solution 2, a stabilizing agent 2, a preservative 2 and a protective agent 2; the polystyrene latex microspheres and the sOB-R antibody are connected by means of covalent cross-linking; the surface functional group of the polystyrene latex microspheres in the reagent R2 is amino, carboxyl, hydrazide, aldehyde or epoxy group. According to the assay kit, a latex reinforced immunonephelometry method is adopted, when the sOB-R in blood reacts with the sOB-R antibody, polystyrene latex is driven to aggregate and generate certain turbidity, so that the detection can be convenient. The sOB-R latex reinforced immunonephelometry assay kit is easily applied in clinical application.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to a sOB-R latex-enhanced immune turbidity detection kit utilizing surface functional groups. Background technique [0002] Soluble leptin receptor (sOB-R) is the most important leptin-binding protein in blood circulation. In the natural state, there are two sizes of leptin-binding protein in human blood, with molecular weights of 140kD and 110kD. After deglycosylation by glycosidase, the molecular weights are 90kD and 60kD. In the human body, soluble leptin receptors mainly come from transmembrane leptin Shedding of the receptor, which has a crucial role in regulating leptin signaling function. sOB-R only exists in the peripheral blood circulation of normal people, and sOB-R has not been found in the cerebrospinal fluid. Under physiological conditions, sOB-R has obvious circadian rhythm, and the level of sOB-R increases when hungry. In healthy people, sOB-R is positiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 潘能科
Owner CHONGQING ZHONGYUAN BIOLOGICAL TECH
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