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Simoa kit of biomarker FOLR1 and using method of Simoa kit

A biomarker and kit technology, applied in the field of immune detection, can solve the problems of high environmental requirements, non-specific reaction effects, easy cross-contamination, etc., and achieve the effects of shortening detection time, reducing manual errors, and wide application range.

Pending Publication Date: 2021-11-05
ZHENGZHOU UNIV
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Problems solved by technology

The instrument uses a flow colorimetric cell, which is easy to cross-contaminate, has extremely high environmental requirements, and is easily affected by environmental factors and other non-specific reactions

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  • Simoa kit of biomarker FOLR1 and using method of Simoa kit

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Embodiment

[0033] The Simoa kit for detecting FOLR1 content in human body fluid provided by the present invention comprises 2.5-2.9 μm magnetic beads coated with capture antibody, FOLR1 standard, biotinylated detection antibody, SBG solution, fluorescent substrate solution and samples Diluents, and electronic files of test methods.

[0034] In the present invention, the raw materials of magnetic beads, capture antibodies, standards, detection antibodies, biotinylation reagents, SBG and fluorescent substrates are purchased; wherein the capture antibody is monoclonal antibody K5646, and the detection antibody is monoclonal antibody K56461 .

[0035] The preparation process of the capture antibody-coated magnetic beads in the present invention is as follows: first, the stored antibody is exchanged through an ultrafiltration tube, and the buffer used is MES, which is 50-60 mmol / mL morpholineethanesulfonic acid buffer, pH=6.2-6.4.

[0036] Then, the magnetic beads were activated with EDC, a...

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Abstract

The invention relates to a single-molecule array detection (Simoa) kit for a biomarker FOLR1 in an immunodetection technology and a using method of the single-molecule array detection (Simoa) kit. The Simoa kit is used for detecting the content of folate receptor 1 (FOLR1) protein in human body fluid based on a Simoa platform single-molecule array technology and a double-antibody sandwich method, and comprises magnetic beads coated with a capture antibody, an FOLR1 standard substance, a biotinylated detection antibody, an SBG solution, a fluorescent substrate solution and a sample diluent. The Simoa kit is advantaged in that sensitivity of the Simoa kit reaches 0.2 pg / mL, the sample loading amount is only 10-20 [mu] L, repeatability is good, the detection range is 0.4-2000 pg / mL, and the content of FOLR1 in human body fluid can be accurately detected.

Description

technical field [0001] The invention relates to the technical field of immunoassay, in particular to a Simoa kit of a biomarker FOLR1 and a use method thereof. Background technique [0002] Folate Receptor 1 (FOLR1) is a membrane protein that is produced in the channel to transport folic acid into cells, and has a high affinity for folic acid and its derivatives. Folic acid plays an important role in regulating cell proliferation, division and tissue growth. The division of tumor cells requires a large amount of folic acid, and the increase of folic acid leads to the upregulation of FOLR1, which is present in ovary, endometrium, lung, breast, high-grade osteosarcoma and pleural mesothelioma. The level of serum FOLR1 increases as the disease progresses and decreases as the body recovers, so it can be used as one of the biomarkers for the clinical diagnosis of lung cancer. [0003] The currently clinically used FOLR1 detection methods are mainly traditional enzyme-linked imm...

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Application Information

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IPC IPC(8): G01N33/577G01N33/68G01N33/574G01N33/543
Inventor 张琳李朝辉谢磊屈凌波
Owner ZHENGZHOU UNIV
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