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.

주의 : 방사성 요오드의 산화 된 형태가 매우 휘발성 적절한 리드 방패와 리드 유리 병 취급해야합니다. 모든 방사 단계는 통풍이 잘되는 숯을 여과 후드에서 수행 한 실험 절차는 방사능 검출 장치에 의해 모니터링해야한다. I 125 - 표지 된 아 지드의 합성 화학 및 역상 카트리지 (1)의 제조 <ol><li> 용액의 시약의 조제 <ol><li> 150 μL의 아 지드 전구체 (2) 무수 에탄올 (그림 1)의 1 mg의 용해. 참고 : 아 지드 전구체에 대한 자세한 합성 절차 (2) 이전의 종이 (22)에보고되었다. </li><li> 1X 인산 완충 생리 식염수 (pH는 7.4) 20 μL를 1 mg을 클로라민 T를 녹인다. </li><li> 20 μL의 H 2 O 2 mg의 나트륨 메타 비설 파이트를 녹여 </li></ol></li><li> Prepara카트리지 기 <ol><li> 10 ㎖의 H 2 O이어서 10 ㎖의 무수 에탄올로 TC18 카트리지 워시 공기와 카트리지의 행렬을 건조하지 마십시오. </li></ol></li></ol> 125 I 표지 아 지드 보철 그룹의 2 Radiosynthesis <ol><li> 전구체의 방사성 요오드화 반응 <ol><li> 1.5 ml의 마이크로 원심 튜브에 지드 전구체 용액 및 아세트산 (10 μL) (무수 에탄올 150 ㎕의 1 mg)을 추가한다. </li><li> 반응 혼합물에 0.1 M의 NaOH (50 μL)에 [125 I]의 NaI 150 MBq의 추가. </li><li> 클로라민 T 용액 (1X 인산 완충 식염수 20 ㎕의 1 mg)을 첨가하고 튜브를 마이크로 원심 함유하는 반응 혼합물을 닫는다. </li><li> 방사성 요오드화 반응이 완료 될 때까지 실온에서 15 분간 반응 혼합물을 인큐베이션. </li><li> 반응 혼합물에 소듐 메타 바이 설 파이트 용액 (2 ㎎을 20 μL의 H 2 O)를 추가방사성 요오드화 반응을 해소합니다. </li><li> 조 생성물을 0.2 μl를 인출 한 후 용액을 100 ㎕로 희석 (H 2 O / CH 3 CN, 1 : 1) HPLC 분석. 참고 : 모든 HPLC 실험의 경우, H 2 O (용매 A)를 포함하는 0.1 % 포름산 및 이동상 0.1 % 포름산 함유 아세토 니트릴 (용매 B)를 사용합니다. </li><li> 칼럼 상 반전 (역상 분석 무선-HPLC를 이용하여 희석하고, 조 생성물을 C18 분석; 유속 : 1 ml / 분 용리액 구배 : 0-2 분 동안 20 % 용매 B, 20 % ~ 80 % 용매 B에 대한 2-22 분, 22 ~ 23 분 80~100% 용매 B, 그리고 23-28 분 동안 100 % 용매 B, 체류 시간 : 16.4 분) (그림 2). </li></ol></li><li> 분취 HPLC로 조 생성물을 정제하여 주 : 이러한 주입기, 컬럼, 검출기 수집 튜브 및 폐수 수집 된 컨테이너 HPLC 부품 주위에 충분한 납 차폐를 제공한다. <ol><li> 일 철수HPLC 바이알에 전체 반응 혼합물을 전자. 아세토 니트릴 (0.5 mL)로 반응 관을 씻어 동일한 주입 병에 린스를 추가합니다. H 2 O (1 mL)로 수집 된 용액을 희석. </li><li> (조제 무선 HPLC 상에 주입하여 조 생성물을 C18 컬럼 상 역방향 유속 : 10 ㎖ / 분 용리액 구배 : 2-22 분 동안 0-2 분, 20 % ~ 80 % 용매 B 20 % 용매 B를 22 ~ 23 분 동안 80~100% 용매 B, 그리고 23-28 분 동안 100 % 용매 B). </li><li> I 125 - 표지 지드 (1)를 나타내는 피크 방사능 수집을 유리 시험관에 (도 2) (이들의 HPLC 조건에서의 t R은 17.8-18.8 분이다). </li><li> 제조사의 프로토콜에 따라 방사선 도즈 교정기를 사용하여 분획의 방사 화학적 수율을 측정한다. </li><li> 제품의 방사 화학적 순도를 결정하기위한 동일한 HPLC 조건을 사용하여 분석 무선 HPLC로 정제 된 생성물을 주입한다. </li></ol></li><li> 제품의 고체상 추출 <ol><li> 40ml의 순수 H 2 O로 원하는 생성물 (1)을 포함하는 분획을 희석 </li><li> 컨디셔닝 TC18 카트리지에 희석액을 추가합니다. </li><li> 추가로 15 ML의 H 2 O와 카트리지를 세척 </li><li> 리드 방패에 의해 보호되는 10 ml의 유리 바이알에 2 ㎖의 아세톤으로 카트리지에 갇혀 제품 (1) 용출. 제조사의 프로토콜에 따라 방사선 도즈 교정을 사용하여 용출 된 생성물의 방사능을 측정한다. 주 : 디메틸 설폭 사이드 (DMSO) 또는 무수 에탄올 또한 카트리지로부터의 생성물의 용출을 위해 사용될 수있다. 방사능의 약 5-10 %는 보통 카트리지 스틱, 남은 방사선 표지 생성물은 완전히 유기 용매의 과잉을 사용하여 용출 될 수 없다. </li><li> 질소 또는 아르곤 가스의 흐름과 함께 아세톤을 증발시켰다. </li><li> 재를 녹여다음 방사성 표지 단계에 대한 DMSO (100-200 μL)와 sidue. </li></ol></li></ol> DBCO 기 - 금 나노 입자 컨쥬 게이트 (3)의 합성 <ol><li> 13 나노 크기의 금 표면 변형 DBCO 기 함유 폴리에틸렌 글리콜 나노 입자 <ol><li> 준비 나트륨 구연산 안정화 된 금 나노 입자 (3) (평균 크기 = 13 ㎚) 이전의 보고서 (24)에 따라. </li><li> (10 nM 내지 15 mL) 중 시트 레이트 - 안정화 된 금 나노 입자에 트윈 20 (1 ㎜, 1.5 ㎖)의 수용액을 추가한다. 궤도 통에 20 분의 솔루션을 흔들어. </li><li> DBCO 기 함유 폴리에틸렌 글리콜 티올 (평균 분자량 = 5,000, 100 μM, 1.5 ㎖)의 수용액을 추가한다. 오비탈 진탕 기에서 2 시간 동안 용액을 흔들어. </li></ol></li><li> DBCO 그룹 - 개질 된 금 나노 입자의 정제 <ol><li> 정제 DBCO 그룹 - 개질 된 금 나노 입자 (4) &lt;/ 연속 원심 분리 (11,400 XG, 15 분 × 3)로&gt; 강한. </li><li> 뜨는을 가만히 따르다 및 금 나노 입자 펠릿의 재 부유 순수을 추가합니다. </li></ol></li></ol> 4. 구리 무료 클릭 반응을 통해 DBCO - 그룹 - 개질 된 금 나노 입자의 방사성 표지 <ol><li> I 125 - 표지 된 아 지드를 사용하여 125 I 표지 된 금 나노 입자의 합성 (1) <ol><li> 원심 분리 (11,400 × g으로 15 분간)을 사용하여 DBCO 그룹 - 개질 된 금 나노 입자의 농축 용액을 제조하고, 2 μM로 금 나노 입자의 농도를 조절한다. </li><li> 금 나노 입자의 현탁액을 DMSO (5 μL)의 125I 표지 지드 (1) 4.1 MBq의 추가 (4) (2 μM, 50 μL). </li><li> 60 분 동안 40 ° C에서 얻어진 반응 혼합물을 인큐베이션. </li><li> 조 생성물로부터 분취 량 (0.2 μL)를 인출하고, 실리카 - 코에 적용ated 박층 크로마토 그래피 (TLC) 판. </li><li> 이동상으로서 에틸 아세테이트를 사용하여 TLC 판을 개발한다. </li><li> 전파 TLC 스캐너의 TLC 플레이트를 배치하고, 제조자의 프로토콜에 따라 방사성 표지 반응 (도 3)을 감시하는 스캐너를 실행. </li></ol></li><li> 조 생성물을 정제하여 <ol><li> 원심 분리하여 125 I 표지 된 금 나노 입자 (4) (11,400 × g으로 15 분간)를 포함하는 반응 혼합물을 정제 하였다. </li><li> 뜨는을 가만히 따르다 및 금 나노 입자 펠릿의 재 부유 순수을 추가합니다. </li><li> 정제 된 제품에서 나누어지는 (0.2 μl를) 철회 및 실리카 코팅 된 TLC 판에 적용합니다. </li><li> 이동상으로서 에틸 아세테이트를 사용하여 TLC 판을 개발한다. </li><li> 무선-TLC 스캐너에 TLC 판을 놓고 125I 표지 GOL의 방사 화학적 수율 및 방사 화학적 순도를 결정하기 위해 스캐너를 실행제조 업체의 프로토콜에 따라 D 나노 입자 (4) (그림 3). </li></ol></li></ol>

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H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+Evaluation+of+an+125I-Labeled+Azide+Prosthetic+Group+for+Efficient+and+Bioorthogonal+Radiolabeling+of+Cyclooctyne-Group+Containing+Molecules+using+Copper-Free+Click+Reaction.">Synthesis and Evaluation of an 125I-Labeled Azide Prosthetic Group for Efficient and Bioorthogonal Radiolabeling of Cyclooctyne-Group Containing Molecules using Copper-Free Click Reaction.</a> Bioorg. Med. Chem. Lett. 26, 875-878 (2016).</li><li>Kim, Y. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+Targeting+and+Imaging+Using+Cyclic+RGD-PEGylated+Gold+Nanoparticle+Probes+with+Directly+Conjugated+Iodine-125.">Tumor Targeting and Imaging Using Cyclic RGD-PEGylated Gold Nanoparticle Probes with Directly Conjugated Iodine-125.</a> Small. 7, 2052-2060 (2011).</li></ol>

The authors have nothing to disclose.

일반적으로, 정제 된 I-125 표지 지드 (1)의 관찰 방사 화학적 수율은 75 ± 10 %이었다 (N = 8). 방사성 표지는 방사능 MBq의 50-150로 수행하고, 그 결과 방사 상당히 일치한다. [125 I]의 NaI (t 1/2 = 59.4 d) 방사성 요오드화 반응에 사용 하였다 개월 이상 방사성 붕괴를 시행 한의 방사 화학적 수율로 관찰하면 약간 (53~65%) 감소 하였다. 따라서, [125</su...

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.

<table border="0" cellpadding="0" cellspacing="0" width="683">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<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.

stannylated 전구체 (2)의 방사성 요오드화 반응은 방사능 표지 된 생성물을 제공하기 위해 실온에서 15 분 동안 [125 I] NaI를 아세트산 및 클로라민 T 150 MBq의를 사용하여 수행 하였다 (1). 조 혼합물을 예비 HPLC 정제 후, 목적 생성물을 방사 화학적 수율은 75 ± 10 % (N = 8)을 얻었다. HPLC 분석은 125 I 표지 된 생성물의 방사 화학적 순도는 99 %...

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

I 125 - 표지 지드의 합성 및 무도 클릭 반응을 이용 dibenzocyclooctyne (DBCO) - 기 - 컨쥬 게이트 13 나노 크기의 금 나노 입자의 방사성 표지에 대한 세부 절차를 설명한다.

를 사용하여 금 나노 입자의 효율적인 방사성 표지를위한 최적화 된 프로토콜<sup&gt; (125)</sup&gt; I-표시 아 지드 보철 그룹

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

an-optimized-protocol-for-efficient-radiolabeling-gold-nanoparticles

Universidad de Cartagena UDEC

Research

JoVE Journal

Chemistry

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

8.1K 조회수.  Korea Atomic Energy Research Institute.  I 125 - 표지 지드의 합성 및 무도 클릭 반응을 이용 dibenzocyclooctyne (DBCO) - 기 - 컨쥬 게이트 13 나노 크기의 금 나노 입자의 방사성 표지에 대한 세부 절차를 설명한다. 

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

Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing and maintaining primary and pluripotent stem cell derived hepatocytes. While biological matrices are useful, they permit short term culture of hepatocytes, limiting their widespread application. We have attempted to overcome the limitations using a synthetic polymer coating. Polymers represent one of the broadest classes of biomaterials and possess a wide range of mechanical, physical and chemical properties, which can be fine-tuned for purpose. Importantly, such materials can be scaled to quality assured standards and display batch-to-batch consistency. This is essential if cells are to be expanded for high through-put screening in the pharmaceutical testing industry or for cellular based therapy. Polyurethanes (PUs) are one group of materials that have shown promise in cell culture. Our recent progress in optimizing a polyurethane coated surface, for long-term culture of human hepatocytes displaying stable phenotype, is presented and discussed.

This article will focus on developing polymer coated surfaces for long-term, stable culture of stem cell derived human hepatocytes.

최적화 된 합성 표면을 간세포 표현형 사용 안정화

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing ...

연구

화학

A Simple Method for the Size Controlled Synthesis of Stable Oligomeric Clusters of Gold Nanoparticles under Ambient Conditions

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like oligomeric clusters of these yellow nanoparticles of narrow size distribution are synthesized under ambient conditions via two methods. The delay-time method controls the number of subunits in the oligoclusters by varying the time between the addition of HAuCl4 to alkaline solution and the subsequent addition of reducing agent, NaSCN. The yellow oligoclusters produced range in size from ~3 to ~25 nm. This size range can be further extended by an add-on method utilizing hydroxylated gold chloride (Na+[Au(OH4-x)Clx]-) to auto-catalytically increase the number of subunits in the as-synthesized oligocluster nanoparticles, providing a total range of 3 nm to 70 nm. The crude oligocluster preparations display narrow size distributions and do not require further fractionation for most purposes. The oligoclusters formed can be concentrated &#62;300 fold without aggregation and the crude reaction mixtures remain stable for weeks without further processing. Because these oligomeric clusters can be concentrated before derivatization they allow expensive derivatizing agents to be used economically. In addition, we present two models by which predictions of particle size can be made with great accuracy.

We describe a simple method for producing highly stable oligomeric clusters of gold nanoparticles via the reduction of chloroauric acid (HAuCl4) with sodium thiocyanate (NaSCN). The oligoclusters have a narrow size distribution and can be produced with a wide range of sizes and surface coats.

주변 조건에 따라 금 나노 입자의 안정 올리고머 클러스터의 크기 제어 합성을위한 간단한 방법

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like ...

A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray coating. These nanoparticles are produced by the polymerization reaction of alkyl cyanoacrylate on the surface of cereal protein (gliadin) molecules. Alkyl cyanoacrylate is a monomer that instantly polymerizes at RT when it is applied to the surface of materials. Its polymerization reaction is initiated by the trace amounts of weakly basic or nucleophilic species on the surface, including moisture. Once polymerized, the polymerized alkyl cyanoacrylates show a strong affinity with the object materials because nitrile groups are in the backbone of poly (alkyl cyanoacrylate). Proteins also work as initiator for this polymerization because they contain amine groups that can initiate the polymerization of cyanoacrylate. If aggregated protein is used as an initiator, protein aggregate is surrounded by the hydrophobic poly(alkyl cyanoacrylate) chains after the polymerization reaction of alkyl cyanoacrylate. By controlling the experimental condition, particles in the nanometer range are produced. The produced nanoparticles readily adsorb to the surface of most materials including glass, metals, plastics, wood, leather, and fabrics. When the surface of a material is sprayed with the produced nanoparticle suspension and rinsed with water, the micellar structure of nanoparticle changes its conformation, and the hydrophilic proteins are exposed to the air. As a result, the nanoparticle-coated surface changes to hydrophilic.

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic. The produced nanoparticle is an assembly of gliadin-cyanoacrylate diblock copolymers. Spray coating with the produced nanoparticle changes the surface of target material to a hydrophilic surface.

친수성 코팅 글 리아 딘 - 시아 노 아크릴 레이트 나노 입자의 생산을위한 프로토콜

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray ...

A Simple and Efficient Protocol for the Catalytic Insertion Polymerization of Functional Norbornenes

Norbornene can be polymerized by a variety of mechanisms, including insertion polymerization whereby the double bond is polymerized and the bicyclic nature of the monomer is conserved. The resulting polymer, polynorbornene, has a very high glass transition temperature, Tg, and interesting optical and electrical properties. However, the polymerization of functional norbornenes by this mechanism is complicated by the fact that the endo substituted norbornene monomer has, in general, a very low reactivity. Furthermore, the separation of the endo substituted monomer from the exo monomer is a tedious task. Here, we present a simple protocol for the polymerization of substituted norbornenes (endo:exo ca. 80:20) bearing either a carboxylic acid or a pendant double bond. The process does not require that both isomers be separated, and proceeds with low catalyst loadings (0.01 to 0.02 mol%). The polymer bearing pendant double bonds can be further transformed in high yield, to afford a polymer bearing pendant epoxy groups. These simple procedures can be applied to prepare polynorbornenes with a variety of functional groups, such as esters, alcohols, imides, double bonds, carboxylic acids, bromo-alkyls, aldehydes and anhydrides.

We describe the catalytic insertion polymerization of 5-norbornene-2-carboxylic acid and 5-vinyl-2-norbornene to form functional polymers with a very high glass transition temperature.

기능 노보 넨의 촉매 삽입 중합을위한 간단하고 효율적인 프로토콜

Norbornene can be polymerized by a variety of mechanisms, including insertion polymerization whereby the double bond is polymerized and the bicyclic ...

A Protocol for Safe Lithiation Reactions Using Organolithium Reagents

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents warrants proper technique, thorough training, and proper personal protective equipment. To aid in the training of researchers using organolithium reagents, a thorough, step-by-step protocol for the safe and effective use of tert-butyllithium on an inert gas line or within a glovebox is described. As a model reaction, preparation of lithium tert-butyl amide by the reaction of tert-butyl amine with one equivalent of tert-butyl lithium is presented.

The safe and proper use of organolithium reagents is described.

유기 리튬 시약을 사용하여 안전 리튬 화의 반응을위한 프로토콜

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents ...

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity and convenient colorimetric readout. However, high densities of nanoparticles are typically needed for detection. The available synthesis-based enhancement protocols are either limited to gold and silver nanoparticles or rely on precise enzymatic control and optimization. Here, we present a protocol to enhance the colorimetric readout of gold, silver, silica, and iron oxide nanoprobes. It was observed that the colorimetric signal can be improved by up to a 10000-fold factor. The basis for such signal enhancement strategies is the chemical reduction of Au3+ to Au0. There are several chemical reactions that enable the reduction of Au3+ to Au0. In the protocol, Good&#39;s buffers and H2O2 are used and it is possible to favor the deposition of Au0 onto the surface of existing nanoprobes, in detriment of the formation of new gold nanoparticles. The protocol consists of the incubation of the microarray with a solution consisting of chloroauric acid and H2O2 in 2-(N-morpholino)ethanesulfonic acid pH 6 buffer following the nanoprobe-based detection assay. The enhancement solution can be applied to paper and glass-based sensors. Moreover, it can be used in commercially available immunoassays as demonstrated by the application of the method to a commercial allergen microarray. The signal development requires less than 5 min of incubation with the enhancement solution and the readout can be assessed by naked eye or low-end image acquisition devices such as a table-top scanner or a digital camera.

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

In this work, a protocol for signal enhancement of nanoprobe-based biosensing is presented. The protocol is based on the reduction of chloroauric acid onto the surface of existing nanoprobes that consist of gold, silver, silica or iron-oxide nanoparticles.

에 라 골드 나노 입자 합성에 의해 급속 한 Nanoprobe 신호 향상

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

The MPLEx Protocol for Multi-omic Analyses of Soil Samples

Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and characterize microbial communities in environmental and biological systems. These measurements are even enabling enhanced analyses of complex soil microbial communities, which are the most complex microbial systems known to date. Multi-omic analyses, however, do have sample preparation challenges, since separate extractions are typically needed for each omic study, thereby greatly amplifying the preparation time and amount of sample required. To address this limitation, a 3-in-1 method for the simultaneous extraction of metabolites, proteins, and lipids (MPLEx) from the same soil sample was created by adapting a solvent-based approach. This MPLEx protocol has proven to be both simple and robust for many sample types, even when utilized for limited quantities of complex soil samples. The MPLEx method also greatly enabled the rapid multi-omic measurements needed to gain a better understanding of the members of each microbial community, while evaluating the changes taking place upon biological and environmental perturbations.

A protocol is presented for simultaneously extracting metabolites, proteins, and lipids from a single soil sample, allowing reduced sample preparation times and enabling multi-omic mass spectrometry analyses of samples with limited quantities.

멀티-omic 토양 샘플의 분석을 위한 MPLEx 프로토콜

Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and ...

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

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 = 640 nm) of BSA-Au(III) complexes. The method adjusts the pH to show that the emergence of the red fluorescence is correlated with the pH-induced equilibrium transitions of the BSA conformations. Red fluorescent BSA-Au(III) complexes can only be formed with an adjustment of pH at or above 9.7, which corresponds to the &#34;A-form&#34; conformation of BSA. The protocol to adjust the BSA to Au molar ratio and to monitor the time-course of the process of Au(III) binding is described. The minimum number of Au(III) per BSA, to produce the red fluorescence, is less than seven. We describe the protocol in steps to illustrate the presence of multiple Au(III) binding sites in BSA. First, by adding copper (Cu(II)) or nickel (Ni(II)) cations followed by Au(III), this method reveals a binding site for Au(III) that is not the red fluorophore. Second, by modifying BSA by thiol capping agents, another nonfluorophore-forming Au(III) binding site is revealed. Third, changing the BSA conformation by cleaving and capping of the disulfide bonds, the possible Au(III) binding site(s) are illustrated. The protocol described, to control the BSA conformations and Au(III) binding, can be generally applied to study the interactions of other proteins and metal cations.

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

The protocols for studying the binding of gold cations (Au(III)) to various conformations of bovine serum albumin (BSA) as well as for characterizing the conformational dependent unique BSA-Au fluorescence are presented.

Gold(III)의 바인딩에 의해 소 혈 청 알 부 민의 다양 한 Conformations에 Luminophore 형성

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

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of their interactions with cell membranes, lipid bilayers, and viruses. The hydrophilic ligands make these particles colloidally stable in aqueous solutions and the combination with hydrophobic ligands creates an amphiphilic particle that can be loaded with hydrophobic drugs, fuse with the lipid membranes, and resist nonspecific protein adsorption. Many of these properties depend on nanoparticle size and the composition of the ligand shell. It is, therefore, crucial to have a reproducible synthetic method and reliable characterization techniques that allow the determination of nanoparticle properties and the ligand shell composition. Here, a one-phase chemical reduction, followed by a thorough purification to synthesize these nanoparticles with diameters below 5 nm, is presented. The ratio between the two ligands on the surface of the nanoparticle can be tuned through their stoichiometric ratio used during synthesis. We demonstrate how various routine techniques, such as transmission electron microscopy (TEM), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), and ultraviolet-visible (UV-Vis) spectrometry, are combined to comprehensively characterize the physicochemical parameters of the nanoparticles.

Amphiphilic gold nanoparticles can be used in many biological applications. A protocol to synthesize gold nanoparticles coated by a binary mixture of ligands and a detailed characterization of these particles is presented.

합성 및 암화 금 나노 입자의 특성화

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

Synthesis of Aptamer-PEI-g-PEG Modified Gold Nanoparticles Loaded with Doxorubicin for Targeted Drug Delivery

Due to drug resistance and toxicity in healthy cells, use of doxorubicin (DOX) has been limited in clinical cancer therapy. This protocol describes the designing of poly(ethylenimine) grafted with polyethylene glycol (PEI-g-PEG) copolymer functionalized gold nanoparticles (AuNPs) with loaded aptamer (AS1411) and DOX through amide reactions. AS1411 is specifically bonded with targeted nucleolin receptors on cancer cells so that DOX targets cancer cells instead of healthy cells. First, PEG is carboxylated, then grafted to branched PEI to obtain a PEI-g-PEG copolymer, which is confirmed by 1H NMR analysis. Next, PEI-g-PEG copolymer coated gold nanoparticles (PEI-g-PEG@AuNPs) are synthesized, and DOX and AS1411 are covalently bonded to AuNPs gradually via amide reactions. The diameter of the prepared AS1411-g-DOX-g-PEI-g-PEG@AuNPs is ~39.9 nm, with a zeta potential of -29.3 mV, indicating that the nanoparticles are stable in water and cell medium. Cell cytotoxicity assays show that the newly designed DOX loaded AuNPs are able to kill cancer cells (A549). This synthesis demonstrates the delicate arrangement of PEI-g-PEG copolymers, aptamers, and DOX on AuNPs that are achieved by sequential amide reactions. Such aptamer-PEI-g-PEG functionalized AuNPs provide a promising platform for targeted drug delivery in cancer therapy.

In this protocol, doxorubicin-loaded AS1411-g-PEI-g-PEG modified gold nanoparticles are synthesized via three-step amide reactions. Then, doxorubicin is loaded and delivered to target cancer cells for cancer therapy.

표적 약물 전달을 위해 독소루비신으로 로드된 압타머-PEI-g-PEG 수정 금 나노 입자의 합성

Due to drug resistance and toxicity in healthy cells, use of doxorubicin (DOX) has been limited in clinical cancer therapy. This protocol describes the ...

를 사용하여 금 나노 입자의 효율적인 방사성 표지를위한 최적화 된 프로토콜

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