11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and application thereof

A hybridoma cell line and monoclonal antibody technology, applied in the field of immunoassay, can solve the problems of high cost, time-consuming, cumbersome steps, etc.

Active Publication Date: 2022-08-02
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] For this reason, the technical problem to be solved by the present invention is to overcome the 11-alpha hydroxyprogesterone content analysis method in the prior art, which takes a long time, has cumbersome steps, and high cost.

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  • 11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and application thereof
  • 11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and application thereof
  • 11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and application thereof

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Embodiment

[0060] A hybridoma cell line that secretes 11-alpha hydroxyprogesterone monoclonal antibody and a preparation method thereof, specifically comprising the following steps:

[0061] (1) Preparation of 11-α hydroxyprogesterone complete antigen:

[0062] a. The derivation route of 11-α hydroxyprogesterone hapten is as follows:

[0063]

[0064] Weigh 10mg of 11-α-hydroxyprogesterone, dissolve it in 0.6mL methanol:water:pyridine (4:1:1) solution, add 4mg carboxymethylhydroxylamine hemihydrochloride (CMO), stir in a water bath at 65°C 2h, let stand overnight at room temperature in the dark. Subsequently, the solution was blown dry by nitrogen, resuspended in 1 mL of chloroform, and extracted three times with ultrapure water. The organic phase was collected, dried with nitrogen again, and the final product was dissolved in 1 mL of anhydrous N,N-dimethylformamide (DMF), namely the 11-α hydroxyprogesterone hapten.

[0065] b. Weigh 7.5 mg of hapten and dissolve it in 500 μL DMF, ...

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Abstract

The invention relates to a 11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and application thereof, and relates to the technical field of immunodetection. The hybridoma cell strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on March 3, 2022, the preservation address is No.3, No.1 yard, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC No.45112. The hybridoma cell strain obtained by the invention belongs to a monoclonal cell strain. The 11-alpha hydroxyprogesterone monoclonal antibody disclosed by the invention has better specificity to 11-alpha hydroxyprogesterone (the cross-over rate to sex hormones such as progesterone, 17-alpha hydroxyprogesterone, medroxyprogesterone, estradiol and the like is less than 0.02%) and detection sensitivity (the IC50 value is 0.311 ng / mL) to the 11-alpha hydroxyprogesterone. And an immunological method is provided for detecting the content of 11-alpha hydroxyprogesterone in a clinical blood sample.

Description

technical field [0001] The invention relates to the technical field of immunodetection, in particular to an 11-alpha hydroxyprogesterone monoclonal antibody hybridoma cell strain and its application. Background technique [0002] Progesterone is a steroid hormone secreted by the corpus luteum of the ovary, also known as progesterone, which is very important as a blood pressure regulator and an anti-androgen. Among them, the progesterone derivatives 11-α- and 11-β-hydroxyprogesterone are potent inhibitors of 11-β-hydroxysteroid dehydrogenase subtype 2 in vitro. To assess whether 11-alpha- and 11-beta-hydroxyprogesterone has a blood pressure-regulating effect, it was found that 14-day infusions of 11-alpha- and 11-beta-hydroxyprogesterone into intact and adrenaltomized Sprague-Dawley rats, 11-alpha- and 11-beta -Hydroxyprogesterone resulted in a significant increase in blood pressure in rats within 3 days, and this effect persisted for 14 days. These results suggest that bot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/26C07K14/765C07K1/107G01N33/74G01N33/577
CPCC07K16/26C07K14/765G01N33/743G01N33/577C07K2317/33
Inventor 胥传来姜晓倩匡华徐丽广孙茂忠吴晓玲刘丽强马伟朱建平郝昌龙宋珊珊胡拥明吴爱红郭玲玲胥欣欣
Owner JIANGNAN UNIV
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