Whole blood-oriented ovarian carcinoma cell detection kit

A detection kit and ovarian cancer cell technology, applied in the fields of biotechnology and biomedicine, can solve the problems of low detection sensitivity and low signal, and achieve the effect of high photochemical stability, high efficiency, and not easy to photolyze

Active Publication Date: 2015-01-28
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Both have their limitations
At present, there are many reports on the preparation of fluorescent immunoprobes by combining QDs with antibodies, but

Method used

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  • Whole blood-oriented ovarian carcinoma cell detection kit
  • Whole blood-oriented ovarian carcinoma cell detection kit

Examples

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Embodiment 1

[0033] Example 1 Ovarian cancer cell detection kit for whole blood detection of ovarian cancer SKOV3 cells

[0034] 1 Preparation of biotinylated BSA-FA

[0035] 1) FA-PEG-NH 2 Coupling: FA 6 mg was dissolved in 600 μL dimethyl sulfoxide, DCC 3 mg, NHSS 3 mg and triethylamine 60 μL were added to react in a dark room for 4 h, and diaminopolyethylene glycol 30 mg was added to react overnight in a dark room ; Centrifuge after the reaction is complete, use acetone at -20°C to precipitate, wash with ethyl acetate three times, dry in a fume hood and store in the dark; 2) Coupling of FA-PEG-BSA: FA-PEG-NH 2 5 mg, EDC 2.5 mg, and NHS 2.5 mg were dissolved in 500 μL of BB at pH 7 for 10 min at room temperature; 8.35 mg of BSA was added, and reacted for 2 h at room temperature, purified by dialysis three times and stored in a refrigerator at 4 °C; 3) Biological Preparation of primed BSA-FA: Mix long-chain biotin: FA-PEG-BSA at a molar ratio of 20:1, incubate at room temperature for 3...

Embodiment 2

[0066] Example 2 Application of Ovarian Cancer Cell Detection Kit for Whole Blood in Whole Blood of Ovarian Cancer Patients

[0067] Take the kit of the present invention: including 1) HE4 goat polyclonal antibody; 2) biotinylated BSA-FA; 3) streptavidinized nano magnetic beads SA-IO; 4) streptavidin The tinylated QDs are SA-QDs; 5) biotinylated anti-HE4 rabbit anti-goat IgG;

[0068] Wherein, each material preparation method is as follows:

[0069] 1 Preparation of biotinylated BSA-FA

[0070] 1) FA-PEG-NH 2 Coupling: FA 6 mg was dissolved in 600 μL dimethyl sulfoxide, DCC 3 mg, NHS 3 mg and triethylamine 60 μL were added to react in a dark room for 4 h, and diaminopolyethylene glycol 30 mg was added to react overnight in a dark room ; Centrifuge after the reaction is complete, use acetone at -20°C to precipitate, wash with ethyl acetate three times, dry in a fume hood and store in the dark; 2) Coupling of FA-PEG-BSA: FA-PEG-NH 2 5 mg, EDC 2.5 mg, and NHS 2.5 mg were dis...

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Abstract

The invention provides a whole blood-oriented ovarian carcinoma cell detection kit. The kit mainly comprises HE4 goat polyclonal antibodies capable of specifically recognizing the antigen of the ovarian carcinoma cell human epididymis protein (HE4), biotinylated bovine serum albumin (BSA)-folic acid (FA), streptavidin-modified nano magnetic beads (SA-IO), streptavidin-modified QDs (SA-QDs), biotinylated anti-HE4 rabbit anti-goat IgG, a connecting reagent and the like. The kit is capable of efficiently separating out the carcinoma cells in the whole blood of an ovarian carcinoma patient by virtue of the enrichment characteristic of the immunomagnetic beads, and realizing multiple-signal synergetic amplification by specially binding with the targeted ovarian carcinoma cells by use of the SA-QDs and a biotinylated HE4 antigen-antibody complex. The kit has high sensitivity and specificity, and has an extremely wide application prospect in early detection of the ovarian carcinoma.

Description

technical field [0001] The invention belongs to the fields of biotechnology and biomedicine. Specifically, the present invention provides a kit for detecting ovarian cancer. Background technique [0002] Ovarian cancer is a common malignant tumor in gynecology. Its incidence rate ranks third among female reproductive system tumors, but its case fatality rate ranks first. Most ovarian cancers are diagnosed at an advanced stage, and their 5-year survival rate is only 30%. Early diagnosis and treatment of ovarian cancer are important factors to improve the prognosis of patients and increase their survival rate. Currently, there are two common clinical screening methods for ovarian cancer: 1) transvaginal ultrasonography; 2) serum CA125 detection. Both have their limitations. Studies have shown that the appearance of circulating tumor cells (CTCs) in the peripheral blood of cancer patients is earlier than that of visible solid tumors. Therefore, finding a method for highly sp...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/531
CPCG01N33/57449
Inventor 许恒毅傅芬聂丽菊叶称连张婉怡李福来
Owner NANCHANG UNIV
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