Double-antibody sandwich enzyme-linked immunoassay method for quantitatively detecting human follicle-stimulating hormone

A sandwich technology of human follicle-stimulating hormone and double antibodies, applied in the field of biomedical enzyme-linked immunoassay, can solve the problems of limited detection range, cumbersome operation, low sensitivity, etc., and achieve stable and repeatable system, wide curve range and high sensitivity Effect

Inactive Publication Date: 2017-12-19
JIANGXI SCI & TECH NORMAL UNIV
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Problems solved by technology

[0006] The present invention provides a double-antibody sandwich enzyme-linked immunoassay for the quantitative detection of human follicle-stimulating hormone (hFSH), which

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  • Double-antibody sandwich enzyme-linked immunoassay method for quantitatively detecting human follicle-stimulating hormone
  • Double-antibody sandwich enzyme-linked immunoassay method for quantitatively detecting human follicle-stimulating hormone
  • Double-antibody sandwich enzyme-linked immunoassay method for quantitatively detecting human follicle-stimulating hormone

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Experimental program
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Effect test

Embodiment 1

[0032] Example 1: Quantitative detection of human urine-derived follicle-stimulating hormone (Lishenbao——quantitative detection system incubated at 37°C for 2 hours)

[0033] (1) Primary antibody coating: Coat 100ul anti-FSH alpha antibody (4ug / ml, i.e. anti-FSH alpha antibody stock (1mg / ml) in 96-well ELISA plate and dilute to 4ug / ml with PBS buffer working solution), overnight at 4°C; the next day, the microtiter plate was washed with PBS buffer solution containing 0.05% Tween-20, that is, washed 4 times with PBST, and washed 3 to 5 times with PBST before each subsequent operation. The cleaning time is 60-90s. At the same time, set 3 groups of controls in the microtiter plate in advance to judge the background effect of the whole system itself, that is, control group 1: use the same volume of PBS buffer solution instead of the standard product; control group 2: only receive FSH beta working solution; control group 3: Just close it.

[0034] (2) Blocking: add 320ul 5% skimm...

Embodiment 2

[0039] Example 2: Quantitative detection of human urine-derived follicle-stimulating hormone (Lishenbao——quantitative detection system incubated at 4°C for 48 hours without shaking)

[0040] (1) Primary antibody coating: Coat 100ul anti-FSH alpha antibody (4ug / ml, i.e. anti-FSH alpha antibody stock (1mg / ml) in 96-well ELISA plate and dilute to 4ug / ml with PBS buffer working solution), overnight at 4°C; the next day, the microtiter plate was washed with PBS buffer solution containing 0.05% Tween-20, that is, washed 4 times with PBST, and washed 3 to 5 times with PBST before each subsequent operation. The cleaning time is 60-90s. At the same time, set 3 groups of controls in the microtiter plate in advance to judge the background effect of the whole system itself, that is, control group 1: use the same volume of PBS buffer solution instead of the standard product; control group 2: only receive FSH beta working solution; control group 3: Just close it.

[0041] (2) Blocking: ad...

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Abstract

The invention provides a double-antibody sandwich enzyme-linked immunoassay method for quantitatively detecting human follicle-stimulating hormone (hFSH). The invention mainly aims to solve the technical problems of low sensitivity, low system stability, limited detection scope, low repeatability, complex operation, and the like, in the prior art. According to the invention, two paired specific monoclonal antibodies (namely, anti-FSH alpha antibody and anti-FSH beta antibody) are adopted for detecting and quantizing human follicle-stimulating hormone or related follicle-stimulating hormone sample. The method provided by the invention has the advantages of easy operation, high sensitivity, high specificity, high system stability, wide FSH detection scope and capability of directly reflecting the concentration or in vitro biologic activity of an FSH standard substance or sample according to a light absorption value (OD value) detected by a microplate reader.

Description

technical field [0001] The invention relates to a double-antibody sandwich ELISA method for quantitatively detecting human follicle-stimulating hormone, and belongs to the technical field of biomedical ELISA. Background technique [0002] Follicle-stimulating hormone (FSH) is a glycoprotein hormone composed of α subunits and β subunits connected by non-covalent bonds. It is one of the main components of drugs commonly used to treat male and female infertility . In women, follicle-stimulating hormone promotes luteinizing hormone production, follicle maturation, and ovulation induction; in men, follicle-stimulating hormone primarily promotes sperm production and maturation. There are currently two types of commercial follicle-stimulating hormone drug preparations on the market, one is urinary-derived follicle-stimulating hormone (urinary-derived FSH, uFSH) extracted from the urine of postmenopausal women, and the other is through DNA recombinant technology The produced recom...

Claims

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Application Information

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IPC IPC(8): G01N33/74
CPCG01N33/74
Inventor 曾斌胡建文刘华马龙华荣茂韩继忠刘梦梦孙云龙
Owner JIANGXI SCI & TECH NORMAL UNIV
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