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Cystatin C detection kit and preparation method thereof

A technology for detecting kits and cystatin, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of increased R&D and production costs, cumbersome reagent preparation process, and low production efficiency, so as to reduce automatic aggregation, Not easy to gather, high precision effect

Inactive Publication Date: 2018-06-01
QINGDAO HIGHTOP BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in most of the preparation methods of cystatin C detection kits, the preparation process of the reagents is relatively cumbersome, requiring multi-step centrifugation, cleaning and ultrasonic resuspension, and the centrifugation speeds of latex microspheres with different particle sizes are different. The ultrasonic resuspension time of the reagent volume is also inconsistent. These steps add some variable factors to the preparation process, and increase the cost of research and development and production. At the same time, the production efficiency is low and the production cycle is long.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A cystatin C detection kit, comprising reagent R1, reagent R2 and cystatin C calibrator, reagent R1 includes preservative 0.8g / L-2g / L, surfactant volume fraction 1%-5%, Emulsifier 0.2g / L~0.5g / L, buffer solution 10mmol / L~200mmol / L; reagent R2 includes cystatin C antibody 40mg / L~45mg / L, polystyrene latex microspheres 10g / L~20g / L L. Latex microsphere activator 25mg / L~36mg / L, sealing agent 50g / L~70g / L, emulsifier 0.2g / L~0.5g / L, preservative 0.8g / L~2g / L, surface active Agent 0.2%~0.5%, buffer solution 10mmol / L~200mmol / L; Cystatin C protein concentration in cystatin C calibrator is 0~8mg / L.

[0036] The buffer comprises at least one of 2-morpholineethanesulfonic acid (MES) buffer, glycine buffer, Goods buffer, borate buffer, phosphate buffer, HEPES buffer, ammonium chloride buffer kind.

[0037]The latex microsphere activator is one of carbodiimide (EDC), N-hydroxysuccinimide (NHS), N-hydroxysulfosuccinimide (Sulfo-NHS) and isocyanate.

[0038] The preservative is one of s...

Embodiment 2

[0045] A preparation method of a cystatin C detection kit comprises a preparation method of reagent R1, a preparation method of reagent R2 and a preparation method of cystatin C calibrator.

[0046] The preparation method of reagent R1 is as follows:

[0047] Weigh 800mL of 50mmol / L glycine buffer solution (pH6.1-6.7), then add 15mL of Tween-20, 0.25g of PEG-20000 and 0.9g of sodium azide, dilute to 1L, and mix well to obtain reagent R1 .

[0048] The preparation method of reagent R2 is as follows:

[0049] (1) Dilute the antibody, dilute the rabbit anti-human cystatin C polyclonal antibody to 1mg / mL with 40mL 50mmol / L MES buffer (pH8.1-8.5) to obtain the diluted rabbit anti-human cystatin C polyclonal antibody solution;

[0050] (2) Dissolve 36 mg of EDC with 4.5 mL of 50 mmol / L MES buffer to obtain an 8 mg / mL activator solution;

[0051] (3) Activation of latex microspheres, adding 4.5mL of activator solution to 20g of polystyrene latex microspheres with a particle size ...

Embodiment 3

[0062] A preparation method of a cystatin C detection kit comprises a preparation method of reagent R1, a preparation method of reagent R2 and a preparation method of cystatin C calibrator.

[0063] The preparation method of reagent R1 is as follows:

[0064] Weigh 800mL of 50mmol / L glycine buffer solution (pH6.1-6.7), then add 50mL of Tween-20, 0.5g of PEG-20000 and 2g of sodium azide, dilute to 1L, and mix well to obtain reagent R1.

[0065] The preparation method of reagent R2 is as follows:

[0066] (1) Dilute the antibody, dilute the rabbit anti-human cystatin C polyclonal antibody to 1mg / mL with 45mL 50mmol / L MES buffer solution (pH8.1-8.5) to obtain the diluted rabbit anti-human cystatin C polyclonal antibody solution;

[0067] (2) Dissolve 25 mg of EDC in 2.5 mL of 50 mmol / L MES buffer to obtain a 10 mg / mL activator solution;

[0068] (3) Activation of latex microspheres, adding 2.5 mL of activator solution to 15 g of polystyrene latex microspheres with a particle s...

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Abstract

The invention provides a cystatin C detection kit and a preparation method thereof. The kit comprises a reagent R1, a reagent R2 and a cystatin C calibration product, wherein the reagent R1 comprisesa preservative, a surfactant, an emulsifier and a buffer solution; the reagent R2 comprises a cystatin C antibody, polystyrene latex microspheres, a latex microsphere activating agent, a sealing agent, an emulsifier, a preservative, a surfactant and a buffer solution. The cystatin C antibody is slowly dropwise added to an activated polystyrene latex microsphere solution, a polystyrene latex microsphere suspension coupled with the cystatin C antibody is obtained after a stirring reaction, sealing is performed, the antibody and latex microspheres are coupled together in a chemical crosslinking manner, and centrifugation, cleaning and ultrasonic resuspension processes in conventional methods are omitted. The production process is simplified, and the kit has the advantages of low blank absorbency, high sensitivity and precision and large linear range.

Description

technical field [0001] The invention belongs to the field of medical in vitro diagnostic reagents, and in particular relates to a cystatin C detection kit and a preparation method thereof. Background technique [0002] The evaluation of glomerular filtration rate (GFR) is very important for the diagnosis of kidney disease. There are many methods to predict GFR, among which the inulin clearance test is the gold standard for evaluating GFR, but this test is complicated to operate and is not suitable for detection in diabetic patients. In addition, inulin can sometimes cause fever symptoms, so it is not often used in clinical diagnosis. On the contrary, it is more convenient and easy to operate to detect the content of creatinine in serum. However, creatinine will be affected by age, gender, nutritional status and diet, so the detection results of different groups of people may be inaccurate. [0003] Cystatin C is the most abundant protease inhibitor in the human body. It is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/68G01N21/31
CPCG01N33/531G01N21/31G01N33/6803G01N2333/8139
Inventor 杨帆于建华程永智徐冬梅宋金玲刘美雪
Owner QINGDAO HIGHTOP BIOTECH
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