Trypsinogen-2 chemiluminescent immunoassay kit and preparation method thereof

A chemiluminescence immunoassay and trypsinogen technology, applied in the field of immunoanalysis medicine, can solve the problems that trypsinogen-2 detection has not yet been applied

Inactive Publication Date: 2011-10-26
BEIJING NYMPHAVN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] As an emerging analytical technique, chemiluminescent immunoassay has not been applied to the detection of trypsinogen-2

Method used

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  • Trypsinogen-2 chemiluminescent immunoassay kit and preparation method thereof
  • Trypsinogen-2 chemiluminescent immunoassay kit and preparation method thereof
  • Trypsinogen-2 chemiluminescent immunoassay kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Application of horseradish peroxidase system to prepare trypsinogen-2 chemiluminescence immunoassay kit of the present invention

[0052] 1. Preparation of horseradish peroxidase-labeled trypsinogen-2 antibody

[0053] Dissolve 5 mg of horseradish peroxidase in 0.5 mL of distilled water, add 0.5 mL of sodium periodate (50 mmol / L) and stir at room temperature for 30 min, dialyze through 10 mM sodium acetate buffer with a pH value of 4.4, add 5 mg of trypsinogen-2 Antibody (Anti-Trypsinogen-28603 from Finland Medix Biochemica Company), stirred for 2h, and finally washed with 200mM NaBH 4 For reduction, after dialysis with 0.02M PBS buffer, add an equal volume of glycerol, and store below -20°C.

[0054] 2. Enzyme-labeled antibody diluent

[0055]

[0056] 3. Enzyme-labeled antibody concentration selection

[0057] The working concentration range of the enzyme-labeled antibody is above 1:500.

[0058] 4. Preparation of trypsinogen-2 antigen calibrator

[...

Embodiment 2

[0082] Example 2 Application of alkaline phosphatase system to prepare pepsin chemiluminescence immunoassay kit of the present invention

[0083] 1. Preparation of alkaline phosphatase-labeled trypsinogen-2 antibody

[0084] Since this kit uses alkaline phosphatase, glutaraldehyde cross-linking method is used to prepare the enzyme label.

[0085] Dissolve 5 mg of alkaline phosphatase in 0.5 mL of distilled water, add 0.5 mL of glutaraldehyde and shake at room temperature for 120 min, dialyze in 10 mM sodium acetate buffer with a pH of 4.4, add 5 mg of trypsinogen-2 antibody (Anti-Trypsinogen-28607) , shake at room temperature for 120 min, and finally remove unlabeled antibodies by polyacrylamide-agarose gel filtration, add an equal volume of glycerol, and store below -20°C.

[0086]Enzyme-labeled antibody diluent

[0087]

[0088]

[0089] Enzyme-labeled antibody concentration selection:

[0090] The working concentration range of the enzyme-labeled antibody is above ...

Embodiment 3

[0098] Example 3 Application of horseradish peroxidase and biotin-streptavidin system to prepare trypsinogen-2 chemiluminescence immunoassay kit of the present invention

[0099] 1. Preparation of biotin-labeled trypsinogen-2 antibody

[0100] (1) Dialyze the trypsinogen-2 antibody (Anti-Trypsinogen-28607) in 1.5mol / L borate buffer solution with pH 8.0 for 12 hours. After the dialysis is completed, take out the antibody and put it into a glass bottle;

[0101] Borate buffer:

[0102]

[0103] (2) Dissolve 800 μg of biotin in DMF;

[0104] (3) Mix the solution obtained in step (1) with the solution obtained in step (2), react at room temperature for 12 hours, and add 100 μl of 2.5 mol / L ammonium chloride solution.

[0105] 2. Preparation of horseradish peroxidase-labeled trypsinogen-2 antibody

[0106] Anti-Trypsinogen-28603 was used to prepare according to the method of Example 1 or 2.

[0107] 3. Preparation of streptavidin-coated solid phase carrier

[0108] (1) Coat...

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Abstract

The invention provides a trypsinogen-2 chemiluminescent immunoassay kit which comprises: 1) a trypsinogen-2 calibrator; 2) a trypsinogen-2 antibody or streptavidin-coated microplate; 3) an enzyme-labelled trypsinogen-2 antibody when a trypsinogen-2 antibody-coated microplate is included in 2), or an enzyme-labelled trypsinogen-2 antibody and a biotin-labelled trypsinogen-2 antibody when a streptavidin-coated microplate is included in 2); 4) a chemiluminescent substrate liquid; and 5) a concentrated washing liquid. The invention also provides a method for preparing the kit. The kit of the invention has the advantages of simple operation, high sensitivity, rapid reaction, etc.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular to a trypsinogen-2 detection kit and a preparation method thereof. Background technique [0002] Acute pancreatitis (AP) is a common disease in abdominal surgery, and the incidence of severe pancreatitis has gradually increased in recent years. Acute pancreatitis causes great disturbance to physiology and obvious damage to important organs, so the mortality rate is very high, and sometimes it can cause sudden death. The mortality rate of severe pancreatitis is 20%, and it can be as high as 50% for those with complications. Therefore, early and accurate diagnosis is needed clinically so as not to delay the disease. [0003] For the diagnosis of acute pancreatitis, amylase is traditionally determined by biochemical methods. However, human amylase has two types of isoenzymes, namely, pancreatic amylase and salivary amylase. The former is specifically secreted by pancreatic cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N21/76G01N33/543G01N33/574
Inventor 陈智周范飞舟范振符张昊岩
Owner BEIJING NYMPHAVN BIOTECH
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