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Large protein pre-S surface antigen for hepatitis B virus chemiluminescence immune assay kit and method for making same

A technology of hepatitis B virus and surface antigen, which is applied in the field of immunoanalysis medicine, can solve the problems of being unable to be widely used in clinical diagnosis and scientific research work, and restricting popularization and use, and achieves the convenience of industrialization and quality control, no cross-reaction, high The effect of sensitivity

Inactive Publication Date: 2009-03-11
北京科美东雅生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In addition, the chemiluminescence immunoassay kit in the prior art is a closed fully automatic chemiluminescence measurement system, which requires an expensive fully automatic chemiluminescence measuring instrument, thus limiting its popularization and use, and cannot be widely used in clinical diagnosis and scientific research.

Method used

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  • Large protein pre-S surface antigen for hepatitis B virus chemiluminescence immune assay kit and method for making same
  • Large protein pre-S surface antigen for hepatitis B virus chemiluminescence immune assay kit and method for making same

Examples

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Embodiment 1

[0045] Example 1 Preparation of the hepatitis B virus surface antigen large protein pre-S region chemiluminescence immunoassay assay kit of the present invention

[0046] 1. Preparation of enzyme-labeled antibody and biotin-conjugated antibody

[0047] 1. The monoclonal antibody to the pre-S region of the large protein of the hepatitis B virus surface antigen is coupled with alkaline phosphatase by the glutaraldehyde method, fully dialyzed against PBS, added with an equal volume of glycerol, and stored below -20°C.

[0048] 2. Preparation of biotin-conjugated monoclonal antibody to the pre-S region of the large protein of hepatitis B virus surface antigen

[0049] (1) Dissolve biotin (BNHS) in N, N-dimethylformamide (DMF) by conventional methods to make 1 mg / mL;

[0050] (2) Use 0.1mol / L NaHCO with a pH value of 9.0 3 Dilute the purified monoclonal antibody to the pre-S region of the large protein of the hepatitis B virus surface antigen to 1-2 mg / mL;

[0051] (3) Mix accor...

Embodiment 2～4

[0099] Embodiments 2-4 Preparation of the hepatitis B virus surface antigen large protein pre-S region chemiluminescence immunoassay assay kit of the present invention

[0100] Except that plastic beads, plastic tubes or magnetic particles are used as carriers respectively, and the pH value of the blocking solution is 7.2, 7.4, 7.6, all the others are prepared in the same way as in Example 1. The pre-S region of the large protein of hepatitis B virus surface antigen Chemiluminescent Immunoassay Assay Kit.

Embodiment 5

[0101] Embodiment 5 The using method of kit of the present invention

[0102] The specific operations of the hepatitis B virus surface antigen large protein pre-S region chemiluminescent immunoassay assay kit prepared in the above embodiment 1 are as follows:

[0103] 1) Preparation of washing buffer: dilute the washing buffer provided in the kit with distilled water 20 times;

[0104] 2) Take the streptavidin-coated solid phase carrier (microwell strip), calibrator / sample to be tested, chemiluminescent substrate solution, and enzyme-labeled antibody out of the refrigerator at 4°C, place it for 15 minutes, and equilibrate to room temperature;

[0105] 3) Fix the microwell plate strip (coated with streptavidin) required for the experiment on the plate holder;

[0106] 4) Add the sample to be tested and 25 μL of 0, 2, 5, 20, 50, and 100 ng / mL to each well of the reaction well, and set a blank well for each test, and then add The mixed volume of biotinylated antibody and enzyme...

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Abstract

The invention relates to the immunoassay medical field, concretely, the invention provides a determination kit of the front S area of the large protein (HBV-LP) of hepatitis B virus surface antigen, and a preparation method thereof. The kit based on the invention comprises: 1) a working calibrator of the front S area of the HBV-LP; 2) a streptavidin carrier; 3) a biotinylation monoclonal antibody of the front S area of the HBV-LP; 4) an alkaline phosphatase-marked monoclonal antibody of the front S area of the HBV-LP; and 5) a chemical luminescent substrate. Further, the preparation method of the kit based on the invention comprises the steps of: 1) preparing the working calibrator by the sterling product of the front S area of the HBV-LP; 2) enveloping the carrier by the streptavidin; 3) carrying out biotinylation to the monoclonal antibody of the front S area of the HBV-LP; 4) marking the monoclonal antibody of the front S area of the HBV-LP by the alkaline phosphatase; 5) preparing the chemical luminescent substrate; 6) subpackaging the working calibrator, an enzyme marker and the chemical luminescent substrate; and 7) assembling and installing for forming finished goods. The kit has the advantages of being simple and convenient, fast, sensitive, stable and the like.

Description

technical field [0001] The present invention relates to the field of immunoanalysis medicine, specifically, the present invention provides a detection kit for the pre-S region of the large protein of hepatitis B virus surface antigen and a preparation method thereof, which combines biotin-avidin immunomagnification technology and chemiluminescence Immunoassay techniques. Background technique [0002] Hepatitis B virus belongs to the family of hepatotropic viruses, which can cause acute hepatitis, chronic hepatitis and liver cirrhosis, and is closely related to the occurrence of liver cancer. China is a high-endemic area of ​​hepatitis B, but there is no effective antiviral method to treat chronic hepatitis B virus infection. Therefore, virus prevention and detection are important means to avoid infection. It provides important information for the development and prognosis of the disease. [0003] HBV surface antigen is the outer membrane protein of HBV, encoded by the S re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/76G01N33/576G01N33/577
Inventor 姚洪涛石根郑金来胡国茂应希堂
Owner 北京科美东雅生物技术有限公司
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