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Detection kit for interleukin-18 (IL-18)

A detection kit and interleukin technology, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problem of low detection sensitivity of interleukin-18 detection

Inactive Publication Date: 2018-04-20
NANTONG EGENS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved by the present invention is the defect that the detection sensitivity of interleukin-18 in the prior art is not high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] This embodiment provides a method for preparing an enzyme conjugate, including the following steps:

[0036] 1. Weigh an appropriate amount of Traut's reagent and use antibody activation buffer (100 mM triethanolamine buffer, pH 8.5) to make a 1.376 mg / mL solution.

[0037] 2. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.

[0038] 3. Store the IL-18 labeled antibody in the antibody activation buffer (100mM triethanolamine buffer, pH8.5) at a concentration of 2mg / mL; add the Traut's solution prepared in step 1 to this solution, and IL-18 labeled The molar ratio of antibody to Traut's reagent is 1:15, mix immediately, and let stand at 25°C for 15 minutes.

[0039] 4. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo-SMCC is 1:10, mix immediately, and statically a...

Embodiment 2

[0050] This embodiment provides a method for preparing an enzyme conjugate, including the following steps:

[0051] 1. Weigh an appropriate amount of Traut's reagent and use antibody activation buffer (100 mM triethanolamine buffer, pH 8.5) to make a 1.376 mg / mL solution.

[0052] 2. Weigh an appropriate amount of Sulfo-SMCC reagent, and use dimethylformamide DMF to prepare a solution with a concentration of 17.5 mg / mL.

[0053] 3. Store the IL-18 labeled antibody in antibody activation buffer (100mM triethanolamine buffer, pH 8.5) at a concentration of 5mg / mL; add Traut's solution prepared in step 1 to this solution, and IL-18 labeled The molar ratio of antibody to Traut's reagent is 1:30, mix immediately, and let stand at 25°C for 15 minutes.

[0054] 4. Add the Sulfo-SMCC solution prepared in step 2 to the alkaline phosphatase solution (AP solution, the concentration is 20mg / ml), the molar ratio of alkaline phosphatase to Sulfo-SMCC is 1:15, mix immediately, and statically at 25°C....

Embodiment 3

[0065] This embodiment provides a method for coating magnetic beads, including the following steps:

[0066] 1. Store the IL-18 coated antibody in 100mM MES buffer pH5.0 at a concentration of 1mg / mL;

[0067] 2. Take a carboxyl magnetic bead solution of 100 times the weight of the antibody, the particle size of the carboxyl magnetic bead is 3μm, magnetically separate and discard the supernatant;

[0068] 3. Washing: Re-dissolve the magnetic beads obtained in the above steps with 100mM MES buffer pH5.0, and magnetically separate and discard the supernatant.

[0069] 4. Repeat step 3 once

[0070] 5. Add an appropriate amount of 100mM MES buffer to dissolve the magnetic beads obtained in step 4.

[0071] 6. Weigh an appropriate amount of NHS reagent and dissolve it into a solution with a concentration of 10 mg / mL in 100 mM MES buffer pH 5.0;

[0072] 7. Weigh an appropriate amount of EDC reagent and dissolve it into a solution with a concentration of 10 mg / mL in 100 mM MES buffer pH 5.0;

[...

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Abstract

The invention discloses a detection kit for IL-18. The detection kit comprises seven storage assemblies which are respectively used for storing an IL-18 calibrator, an IL-18 control material, an enzyme conjugate working solution, a magnetic bead working solution, a rinsing solution, a substrate solution and a pretreatment reagent, wherein the magnetic bead working solution comprises carboxyl magnetic beads labeled by an IL-18 antibody, and the enzyme conjugate working solution comprises an alkaline phosphatase-labeled anti-IL-18 antibody. The invention also discloses a detection method for detecting IL-18. The detection kit disclosed in the invention has a minimum detection limit of 5 pg / ml, a linear range of 5-5000 pg / ml, high detection sensitivity and a wide linear range, enables detection time to be shortened to 15 min and simplifies detection steps.

Description

Technical field [0001] The invention relates to the field of biological detection, in particular to an interleukin-18 detection kit. Background technique [0002] Acute kidney injury (Acute kidney injury, AKI) is a complex hyporenal syndrome that is caused by many reasons and can occur in various clinical situations. It can be manifested as a slight increase in blood creatinine levels or as acute renal failure. Studies have found that the occurrence of AKI is closely related to the increase in mortality. This requires physicians to make early diagnosis in clinical work. It is best to detect and intervene in time when the glomerular filtration rate begins to decrease, or when there is only tissue damage and the glomerular filtration rate is still normal, to improve the prognosis and reduce Case fatality rate. Serum creatinine and urine output are the current diagnostic indicators of AKI, but they are affected by many factors, and their sensitivity and specificity are not high. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/543
CPCG01N33/535G01N33/54326
Inventor 王保君欧卫军沙海苏玲玲
Owner NANTONG EGENS BIOTECH CO LTD
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