Preparation method and application of avian leukemia virus antigen immunosensor
A technology for avian leukosis virus and antigen immunization, which is applied in the field of immune analysis and biosensing, can solve the problems of time-consuming, etc., and achieve the effects of less reagent consumption, simple preparation, and easy miniaturization
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Embodiment 1
[0027] Example 1 Core-shell nanomaterial amino-β-CDFe 3 o 4 SiO 2 - Preparation of Fc
[0028] 2.5g of Fe 3 o 4 ·7H 2 O with 4.0g Fe 2 (SO 4 ) 3Dissolve in a 500mL flask filled with 200mL of ultrapure water, add 50mL of ammonia water with a mass fraction of 25% to adjust the pH to 10, stir at a constant temperature of 45 °C for 1 h, and cool to room temperature; add 8 mL of tetraethyl ortho silicon salt TEOS, stirred for 16 h, centrifuged, and vacuum-dried at 50°C for 12 h to obtain Fe 3 o 4 SiO 2 ;
[0029] Take 0.08g Fe 3 o 4 SiO 2 Mix with 0.1 mL of aminopropyltriethoxysilane APTES and 10 mL of toluene, reflux at 110 °C for 1.5 h, and magnetically separate with a strong magnet. The obtained product is washed with methanol and ethanol three times in turn, and dried in vacuum for 12 h to obtain Aminated Fe 3 o 4 SiO 2 ;
[0030] 40 mg of aminated Fe 3 o 4 SiO 2 Disperse into 1 mL of absolute ethanol, add 250-350 μL of glutaraldehyde with a mass fraction ...
Embodiment 2
[0031] Example 2 Core-shell nanomaterial amino-β-CDFe 3 o 4 SiO 2 - Preparation of Fc
[0032] 3.5g of Fe 3 o 4 ·7H 2 O with 4.0g Fe 2 (SO 4 ) 3 Dissolve in a 500mL flask filled with 200mL of ultrapure water, add 50mL of ammonia water with a mass fraction of 28% to adjust the pH to 10, stir at a constant temperature of 55°C for 1 h, and cool to room temperature; add 12 mL of tetraethyl ortho silicon salt TEOS, stirred for 20 h, centrifuged, and vacuum-dried at 50°C for 12 h to obtain Fe 3 o 4 SiO 2 ;
[0033] Take 0.12g Fe 3 o 4 SiO 2 Mix with 0.1 mL of aminopropyltriethoxysilane APTES and 10 mL of toluene, reflux at 110 °C for 1.5 h, and perform magnetic separation with a strong magnet. The obtained product is washed with methanol and ethanol four times in turn, and dried in vacuum for 12 h to obtain Aminated Fe 3 o 4 SiO 2 ;
[0034] 60 mg of aminated Fe 3 o 4 SiO 2 Disperse into 1 mL of absolute ethanol, add 350 μL of glutaraldehyde with a mass fractio...
Embodiment 3
[0035] Example 3 Amination β -CDFe 3 o 4 SiO 2 -Fc / Ab 2 preparation
[0036] Aminated 3.5 mg β -CDFe 3 o 4 SiO 2 -Fc was dispersed in 1 mL of PBS solution with pH 7.0, sonicated, and aminated β -CDFe 3 o 4 SiO 2 -Fc suspension, add 10 mg of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride EDC and 10 mg of N-hydroxysuccinimide NHS, shake at room temperature for 15 min, Add 0.5 mL, 80 μg·mL -1 detection antibody Ab 2 The solution was incubated at 4°C with shaking for 12 h, centrifuged to discard the supernatant, washed and separated with PBS solution with pH=7.0, added 200 μL of PBS solution, and placed in a refrigerator at 4°C for use.
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