Ovarian Cancer Cell Detection Kit for Whole Blood
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 effects of high photochemical stability, high efficiency and fast speed
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Embodiment 1
[0032] Example 1 Ovarian cancer cell detection kit for detection of ovarian cancer SKOV3 cells in whole blood
[0033] 1 Preparation of biotinylated BSA-FA
[0034] 1) FA-PEG-NH 2 Coupling: Dissolve 6 mg of FA in 600 μL dimethyl sulfoxide, add 3 mg of DCC, 3 mg of NHSS and 60 μL of triethylamine to react in a dark room for 4 hours, add 30 mg of diaminopolyethylene glycol and react overnight in a dark room; after the reaction is complete, centrifuge and store at -20 °C Precipitate with acetone, 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 Dissolve 5mg, EDC2.5mg, NHS2.5mg in 500μL BB with pH 7 and react together at room temperature for 10min; add BSA8.35mg, react at room temperature for 2h, dialyze and purify for 3 times, and store in refrigerator at 4℃; 3) Biotinylated BSA-FA Preparation: mix long-chain biotin: FA-PEG-BSA at a molar ratio of 20:1, incubate at room temperature for 30 min, dialyze the mixe...
Embodiment 2
[0065] Example 2 Application of Ovarian Cancer Cell Detection Kit for Whole Blood in Whole Blood of Ovarian Cancer Patients
[0066] 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;
[0067] Wherein, each material preparation method is as follows:
[0068] 1 Preparation of biotinylated BSA-FA
[0069] 1) FA-PEG-NH 2 Coupling: FA6mg was dissolved in 600μL dimethyl sulfoxide, DCC3mg, NHS3mg and triethylamine 60μL were added to react in a darkroom for 4 hours, and diaminopolyethylene glycol 30mg was added to react overnight in a darkroom; after the reaction was complete, centrifuge and store at -20℃ Precipitate with acetone, 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 Dissolve 5mg, EDC2.5mg, NHS2.5mg in 500μL ...
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