Estriol-specificity-resistant monoclonal antibody hybridoma cell strain NaN-2 and application thereof

A hybridoma cell line, monoclonal antibody technology, applied in specific peptides, biochemical equipment and methods, material testing products, etc., can solve the problems of expensive instruments, short shelf life of markers, restricting the promotion of detection methods, etc. The effect of simplified steps, good detection sensitivity and affinity

Active Publication Date: 2016-08-10
深圳重链生物科技有限公司
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Problems solved by technology

Clinically, radioimmunoassay is mainly used to detect estriol. This method has the disadvantages of radioisotope waste disposal and short shelf life of markers.
The above detection methods can be used for quantitative analysis and have a low detection limit, but usually require expensive instruments and complicated operations, and the pretreatment and detection time is long, which seriously restricts the promotion of these detection methods

Method used

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  • Estriol-specificity-resistant monoclonal antibody hybridoma cell strain NaN-2 and application thereof
  • Estriol-specificity-resistant monoclonal antibody hybridoma cell strain NaN-2 and application thereof
  • Estriol-specificity-resistant monoclonal antibody hybridoma cell strain NaN-2 and application thereof

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Embodiment 1

[0019] Embodiment 1 Preparation of hybridoma cell line NaN-2

[0020] (1) Synthesis of complete antigen: 300mg (1.10 mmol) E 3 Dissolve in 6 mL of dry dimethyl sulfoxide and add 1 g of KOH powder. After stirring for 5 min, 300 mg of bromohexanoic acid was added. Stirring was continued, and 50 mL of ice water was added after 2 h of reaction. Ethyl acetate extraction recovers unreacted E 3 . The aqueous phase was acidified with 2mol / L HCl, and a white precipitate appeared. The sand core funnel was filtered, and the precipitate was washed with distilled water until neutral, and dried in vacuum. Methanol-chloroform recrystallization gave colorless crystals, namely the estriol hapten. Take 4.5mg of the above hapten, then add 2.0mg EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) and 1.0mg NHS (N-hydroxysuccinimide) , dissolved in DMF (N,N-dimethylformamide), stirred at room temperature, and activated for 4 hours; another 5 mg of BSA (bovine serum albumin) wa...

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Abstract

The invention provides an estriol-specificity-resistant monoclonal antibody hybridoma cell strain NaN-2 and application thereof and belongs to the technical field of food safety immune detection. The estriol-specificity-resistant monoclonal antibody hybridoma cell strain NaN-2 is preserved in General Microbiology Center, Microbial Preservation Management Committee, China, and the preservation number is CGMCC No. 12016. An estriol-specificity-resistant monoclonal antibody is generated by secretion of the hybridoma cell strain NaN-2. The monoclonal antibody secreted by the cell strain has good specificity to estriol (the IC 50 value is 0.25 ng / mL) and can be used for estriol specificity detection in food safety. The obtained estriol-resistant monoclonal antibody cell strain has good detection sensitivity and appetency to estriol, a novel method for synthesizing estriol immunogen is further provided, the synthesis step of hapten is more simplified and effective, and a thought and method for synthesizing immunogen are provided for people to conduct research in the future.

Description

technical field [0001] The invention relates to an anti-estriol-specific monoclonal antibody hybridoma cell line NaN-2 and an application thereof, belonging to the technical field of food safety immunoassay. Background technique [0002] Estriol (E 3 ) is an endogenous steroid hormone that is estradiol (E 2 ), estrone (E 1 ) metabolites, E in pregnant women 3 It is mainly derived from the placenta, which is an important indicator of fetal development and placental function in late pregnancy, and is also a common indicator of estrogen in cosmetics. Estriol is used in animal husbandry to promote the growth and development of livestock. Improper use of estriol may lead to residual pollution of estriol in aquatic products, meat, eggs and milk. At present, my country has explicitly prohibited the addition of estriol in the feeding of food animals. [0003] At present, the detection of estriol mostly adopts high performance liquid chromatography, liquid chromatography and mas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44G01N33/74G01N33/577C12R1/91
CPCC07K16/44C07K2317/92G01N33/577G01N33/74
Inventor 刘丽强王忠兴胥传来匡华徐丽广马伟吴晓玲宋珊珊胡拥明
Owner 深圳重链生物科技有限公司
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