GNPs–PEG@FITC 的制备及其在荧光成像中的应用-UDP糖丨MOF丨金属有机框架丨聚集诱导发光丨荧光标记推荐西安齐岳生物

Preparation of FITC coupled GNPs–PEG@cNGR and GNPs–PEG.To prepare FITC coupled GNPs–PEG@cNGR and GNPs–PEG, 1 mg of the GNPs–PEG@cNGR probe, previously synthesized, was dispersed into 1 mL crosslinking reaction solution containing 7.56 mg Na2CO3, 1.06 mg NaHCO3, and 7.36 mg NaCl, sonicated for 2 min, and stored at 4 °C for 10 min. Subsequently, 0.3 mg FITC in 300 μL of DMSO was poured into the above solution under gentle and continuous stirring and incubated in the dark for 8 h at 4 °C. Next, 20 μL NH4Cl solution (5 M) was poured into the above solution under gentle and continuous stirring to terminate the above reaction and then incubated in the dark for 2 h at 4 °C. The final GNPs–PEG@cNGR@FITC was retrieved by ultrafiltration with a Millipore centrifugal filter unit (Amicon Ultra-0.5; MWCO 100[thin space (1/6-em)]000), washed three times, and resuspended in 0.5 mL of PBS (pH 7.4).

In addition, FITC-conjugated GNPs–PEG (GNPs–PEG@FITC) was also prepared as a control. Briefly, the preparation was similar to the fabrication of GNPs–PEG@cNGR, but 220 μL SH–cNGR (1 mg mL−1) was replaced by 15 μL 2-aminoethanethiol (20 mM).

GNPs–PEG@FITC

准备FITC耦合的GNP——PEG@cNGR以及GNP-PEG，1毫克GNP-PEGPEG@cNGR将之前合成的探针分散到含有7.56mg Na2CO3、1.06mg NaHCO3和7.36mg NaCl的1mL交联反应溶液中，超声处理2分钟，在4°C下储存10分钟。

随后，在温和连续搅拌下，将0.3mg FITC溶于300μL DMSO中倒入上述溶液中，在4℃下避光孵育8小时。接下来，在温和连续搅拌下将20μL NH4Cl溶液（5M）倒入上述溶液中以终止上述反应，然后在4°C下在黑暗中孵育2小时。最终的GNPPEG@cNGR@使用Millipore离心过滤器单元（Amicon Ultra-0.5；MWCO 100[薄空间（1/6-em）]000）进行超滤，回收FITC，洗涤三次，并重新悬浮在0.5 mL PBS（pH 7.4）中。

此外，FITC偶联的GNP-PEG（GNP-PEG）PEG@FITC)也准备作为对照。简而言之，制备过程类似于GNP的制造——PEG@cNGR，但220μL SH-cNGR（1 mg mL-1）被15μL 2-氨基乙硫醇（20 mM）所取代。