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Ion stabilizing agent and suspension stabilizing agent cooperatively used kit for detecting sOB-R (soluble leptin receptor) by latex enhanced turbidimetric immunoassay

A detection kit and suspension stabilizer technology, applied in the field of sOB-R latex-enhanced immune turbidimetric detection kits, can solve the problems of poor control of sample dilution, narrow linear range, complicated operation, etc. The effect of increasing sensitivity and extending the validity period

Active Publication Date: 2015-02-18
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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  • Ion stabilizing agent and suspension stabilizing agent cooperatively used kit for detecting sOB-R (soluble leptin receptor) by latex enhanced turbidimetric immunoassay

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

[0062] Preparation of R1 reagent:

[0063] 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.

[0064] Preparation of R2 reagent:

[0065] 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.

[0066] 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 in particular relates to an ion stabilizing agent and suspension stabilizing agent cooperatively used kit for detecting a soluble leptin receptor (sOB-R) by latex enhanced turbidimetric immunoassay. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is formed by a buffer solution 1, a stabilizing agent 1, a preservative 1, ethylene diamine tetraacetic acid (EDTA), a coagulant and a protective agent 1; the reagent R2 is formed by polystyrene latex microspheres crosslinked with an sOB-R antibody, buffer solutions 2, stabilizing agents 2, a preservative 2 and a protective agent 2; the polystyrene latex microspheres are connected with the sOB-R antibody by covalent crosslinking; the stabilizing agents 2 in the reagent R2 are an ion stabilizing agent and a suspension stabilizing agent, which are cooperatively used. The kit has the beneficial effects that latex enhanced turbidimetric immunoassay is adopted, and when the sOB-R in the blood reacts with the sOB-R antibody, polystyrene latexes are driven to gather to generate a certain turbidity, so that the kit is more convenient in detection and is easy to apply clinically.

Description

technical field [0001] The invention relates to the field of in vitro diagnostic reagents, in particular to an sOB-R latex-enhanced immune turbidimetric detection kit which is used in conjunction with an ion stabilizer and a suspension stabilizer. 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 ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/531G01N33/54313
Inventor 吴勇泉
Owner CHONGQING ZHONGYUAN BIOLOGICAL TECH
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