CDs/SiO2-SFTSV monoclonal antibody conjugate and preparation method and application thereof
A monoclonal antibody and polyclonal antibody technology, applied in the field of biochemical analysis and detection, can solve the problems of cumbersome detection technology and low detection limit, and achieve the effects of high fluorescence intensity, improved detection limit and strong specificity
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Embodiment 1
[0040] The preparation of embodiment 1 silanized CDs
[0041] 1) Preparation of reactants before preparation
[0042] H in citric acid monohydrate 2 O molecules interfered greatly with the synthesized silylated CDs, so citric acid monohydrate was dried overnight in a high-temperature vacuum oven for dehydration before the experimental reaction.
[0043] 2) Preparation of silylated CDs
[0044] Pour 15mL of AEAPTMS into a 50mL three-necked flask and blow it with nitrogen for 15min. The solution was then heated to 230°C and 1 g of dehydrated citric acid monohydrate was added rapidly with vigorous stirring. After 5 min, the heating was stopped, and the CDs solution was obtained by natural cooling.
[0045] 3) Purification of silanized CDs
[0046] The obtained CDs solution was centrifuged at 10,000 rpm for 10 min to remove large particles produced by the reaction for purification to obtain CDs.
Embodiment 2
[0047] Example 2 CDs / SiO 2 Preparation of fluorescent nanospheres
[0048] 1) CDs / SiO2 2 Preparation of fluorescent nanospheres
[0049] In 99mL water, add 0.2g CTAB and 60mg NaOH, and stir at 80°C for 1h, where the final concentrations of CTAB and NaOH are 2mg / ml and 0.6mg / ml respectively; then continue to add 1.2mLTEOS and 0.35mL under strong stirring to implement The mixed solution of CDs prepared in Example 1 was reacted for 3h, and the precipitate was collected by centrifugation, and was washed with ethanol and H 2 O washed several times to obtain the reaction product. The final volume ratios of TEOS and CDs in water are 1.2% and 0.35%, respectively.
[0050] 2) CDs / SiO2 2 Washing and purification of fluorescent nanospheres
[0051] The reaction product was dispersed in a mixture of 15 mL HCl and 120 mL ethanol and stirred at 60° C. for 6 h to extract the residual organic template. And repeat the extraction once, the product was washed to neutral, and finally the o...
Embodiment 3
[0052] Example 3 CDs / SiO 2 - Preparation of SFTSV monoclonal antibody conjugates
[0053] 1) CDs / SiO2 2 Carboxylation of fluorescent nanospheres
[0054] The amino-terminated CDs / SiO prepared in Example 2 2 The sphere solution was centrifugally dispersed in DMF, CDs / SiO 2 The concentration of spheres in DMF solution was 2 mg / mL. Then add succinic anhydride, the concentration of succinic anhydride in DMF solution is 5mg / mL, CD / SiO 2 The spheres were carboxylated and stirred for 4 hours. Carboxy-terminated CD / SiO 2 The spheres were washed several times with ethanol and water and dispersed in 2-(N-morpholino)ethanesulfonic acid buffered saline (MES, 0.01M, pH=6.0) for further use. CDs / SiO2 2 Fluorescent nanospheres and CDs / SiO 2 The ZeTa potential in aqueous solution after carboxylation of fluorescent nanospheres is as follows: Figure 4 .
[0055] 2) CDs / SiO2 2 - Preparation of SFTSV monoclonal antibody conjugates
[0056] Functionalization of carboxy-terminated CDs...
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