Name: an optimized protocol for the efficient radiolabeling of gold nanoparticles by using a 125i-labeled azide prosthetic group
Uploaded: 2016-10-10T15:00:00
Description: Watch this Scientific Journal Video about An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group at JoVE.com

This work was supported by grants from the National Research Foundation of Korea, funded by the government of the Republic of Korea, (Grant nos. 2012M2B2B1055245 and 2012M2A2A6011335) and by the RI-Biomics Center of Korea Atomic Energy Research Institute.

注意：放射性碘的氧化形式是相当挥发性的，且必须有足够的铅屏蔽和铅的小瓶处理。所有放射化学步骤应在通风良好的木炭过滤通风橱中进行，实验程序需要通过放射性检测装置进行监控。 1.化学品和反向相萃取柱的制备125 I标记的叠氮合成<ol><li> 在溶液试剂的制备 <ol><li>溶解1毫克150微升叠氮化物前体（2）无水乙醇（ 图1）。 注：叠氮...

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在一般情况下，纯化的125 I标记的叠氮化物（1）所观察到的放射化学产率为75±10％（N = 8）。放射性标记用放射性50-150活度完成，放射化学的结果是相当一致的。如果[125 I]碘化钠（T 1/2 = 59.4 D）将经历放射性衰变了一个多月的放射性碘化反应时，观察到1放化收益率将略有下降（53-65％）。因此，我们建议，因为它产生的[125 I]碘化钠?...

The strain-promoted copper-free click reaction between azides and cyclooctynes has been extensively applied to the efficient bioorthogonal labeling of a wide range of biomolecules, nanomaterials, and living subjects1-7. Due to the excellent site-specificity and rapid reaction rate of this conjugation reaction, it has also been used to synthesize radiolabeled tracers. A few 18F-labeled azide or DBCO prosthetic groups have been prepared for in vitro labeling of various cancers targeting peptides and antibodies, as well as for in vivo pre-targeted imaging of tumors8-13. In addition to these examples, the same conjugation reaction was applied to the metal-radioisotope-labeling of nanomaterials for positron emission tomography (PET) imaging studies14-16.
For several decades, radioactive iodines have been used for biomedical research and clinical trials through PET imaging (124I), single-photon emission computed tomography (SPECT) imaging (123I, 125I), and thyroid cancer treatment (131I)17-21. Therefore, an efficient method for radioactive iodine labeling is fundamentally important for various investigations, including molecular imaging studies, analysis of organ distribution of biomolecules, biomarker identification, and drug development. A copper-free click reaction strategy could be used in radioactive iodine labeling. However, this application has not been investigated as extensively as 18F-labeled biomolecules22-23. Here, we will provide a step-by-step protocol for the synthesis of an 125I-labeled azide for radiolabeling of DBCO-group-derived molecules. The procedures in the present report will include radioiodination of the stannylated precursor, purification steps with HPLC, and solid phase extraction. We also demonstrate efficient radiolabeling of DBCO-group-modified 13-nm-sized gold nanoparticles using the 125I-labeled azide. The detailed protocol in this report will help synthetic chemists understand a new radiolabeling methodology for the synthesis of radiolabeled products.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Chloramine T trihydrate</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">402869</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:255px;">[125I]NaI in aq. NaOH</td>
			<td style="width:101px;">Perkin-Elmer</td>
			<td style="width:160px;">NEZ033A010MC</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Sodium metabisulfite&#160;</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">S9000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Formic acid</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">251364</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Sep-Pak tC18 plus cartridge</td>
			<td style="width:101px;">Waters</td>
			<td>WAT036800</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Dimethyl sulfoxide&#160;</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">D2650</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Acetone</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">650501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Ethanol</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">459844</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Gold(III) chloride trihydrate</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">520918</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Tween 20&#160;</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">P1379</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">DBCO PEG SH (MW 5000)</td>
			<td style="width:101px;">NANOCS</td>
			<td style="width:160px;">PG2-DBTH-5k</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:255px;">TLC silica gel 60 F254</td>
			<td style="width:101px;">Merck</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Analytical HPLC</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">1290 Infinity</td>
			<td>Model number</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Preparative HPLC</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">1260 Infinity</td>
			<td>Model number</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">Analytical C18 reverse-phase column</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">Zorbax Eclipse XDB-C18</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Preparative C18 reverse-phase column</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">PrepHT XDB-C18</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Radio TLC scanner</td>
			<td style="width:101px;">Bioscan</td>
			<td style="width:160px;">AR-2000</td>
			<td>Model number</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Radioisotope dose calibrator</td>
			<td style="width:101px;">Capintec, Inc</td>
			<td style="width:160px;">CRC -25R dose calibrator</td>
			<td>Model number</td>
		</tr>
	</tbody>
</table>

an optimized protocol for the efficient radiolabeling of gold nanoparticles by using a 125i-labeled azide prosthetic group

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient radiolabeling of DBCO-group-functionalized gold nanoparticles using a copper-free click reaction. Radioiodination of the stannylated precursor (2) was carried out by using [125I]NaI and chloramine T as an oxidant at room temperature for 15 min. After HPLC purification of the crude product, the purified 125I-labeled azide (1) was obtained with high radiochemical yield (75 &plusmn; 10%, n = 8) and excellent radiochemical purity (&gt;99%). For the synthesis of radiolabeled 13-nm-sized gold nanoparticles, the DBCO-functionalized gold nanoparticles (3) were prepared by using a thiolated polyethylene glycol polymer. A copper-free click reaction between 1 and 3 gave the 125I-labeled gold nanoparticles (4) with more than 95% of radiochemical yield as determined by radio-thin-layer chromatography (radio-TLC). These results clearly indicate that the present radiolabeling method using a strain-promoted copper-free click reaction will be useful for the efficient and convenient radiolabeling of DBCO-group-containing nanomaterials.

所述stannylated前体（2）的放射性碘化反应进行了使用[125 I]碘化钠，乙酸，和氯胺T 150活度在室温下15分钟，以提供放射性标记的产物（1）。粗混合物的制备HPLC纯化后，用放射化学产率的75±10％（N = 8）中得到所需产物。分析性HPLC显示，125 I标记产物的放射化学纯度是99％以上（ 图2），和产品1所观察到的?...

Watch this Scientific Journal Video about An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group at JoVE.com

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

对于一个125 I标记的叠氮化物的合成和dibenzocyclooctyne（DBCO） -基共轭，13纳米大小的金使用无铜点击反应的纳米粒子的放射性标记的详细过程进行说明。

一个优化的协议通过使用金纳米粒子的放射性标记效率<sup&gt; 125</sup&gt;我标记的叠氮辅基

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient ...

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An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

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Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

原位金纳米粒子合成快速 Nanoprobe 信号增强

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity ...

Luminophore Formation in Various Conformations of Bovine Serum Albumin by Binding of Gold(III)

Luminophore Formation in Various Conformations of Bovine Serum Albumin by Binding of GoldIII

结合金 (III.) 对牛血清白蛋白不同构象的 Luminophore 形成

The purpose of the presented protocols is to study the process of Au(III) binding to BSA, yielding conformation change-induced red fluorescence (&#955;em ...

Synthesis and Characterization of Amphiphilic Gold Nanoparticles

两栖金纳米粒子的合成与表征

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of ...