文獻(xiàn)：Tumor angiogenesis targeting and imaging using gold nanoparticle probe with directly conjugated cyclic NGR?

作者：Minghao Wu? ORCID logoa, Yanyan Zhang?a, Ying Zhanga, Mingjie Wub, Menglin Wua, Hongyi Wua, Lin Caoa, Liang Lia, Xue Li*a and Xuening Zhang*a

文獻(xiàn)鏈接：https://pubs.rsc.org/en/content/articlehtml/2018/ra/c7ra10155d

摘要：

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).

準(zhǔn)備FITC耦合的GNP——PEG@cNGR以及GNP-PEG，1毫克GNP-PEGPEG@cNGR將之前合成的探針?lè)稚⒌胶��?.56mg Na2CO3、1.06mg NaHCO3和7.36mg NaCl的1mL交聯(lián)反應(yīng)溶液中，超聲處理2分鐘，在4°C下儲(chǔ)存10分鐘。

隨后，在溫和連續(xù)攪拌下，將0.3mg FITC溶于300μL DMSO中倒入上述溶液中，在4℃下避光孵育8小時(shí)。接下來(lái)，在溫和連續(xù)攪拌下將20μL NH4Cl溶液（5M）倒入上述溶液中以終止上述反應(yīng)，然后在4°C下在黑暗中孵育2小時(shí)。最終的GNPPEG@cNGR@使用Millipore離心過(guò)濾器單元（Amicon Ultra-0.5；MWCO 100[薄空間（1/6-em）]000）進(jìn)行超濾，回收FITC，洗滌三次，并重新懸浮在0.5 mL PBS（pH 7.4）中。

此外，F(xiàn)ITC偶聯(lián)的GNP-PEG（GNP-PEG）PEG@FITC)也準(zhǔn)備作為對(duì)照。簡(jiǎn)而言之，制備過(guò)程類似于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。

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