이 연구는 중국의 국립 자연 과학 재단에 의해 투자되었다 (31700840); 허난성의 주요 과학 연구 프로젝트 (18B430013, 18A150049). 이 연구는 XYNU의 젊은 학자를위한 난후 학자 프로그램에 의해 지원되었다. 저자는 그의 도움이 작품에 대한 XYNU생명과학대학의 학사 학생 제보 Qu에게 감사를 표하고 싶습니다. 저자는 장비 사용에 대한 XYNU의 분석 및 테스트 센터를 인정하고 싶습니다.

주의: 모든 화학 물질을 사용하기 전에 모든 관련 재료 안전 데이터 시트(MSDS)를 참조해야 합니다. 중합체 및 나노 입자를 준비하는 데 사용되는 화학 물질의 몇 가지는 급성 독성이 있습니다. 나노 입자는 또한 잠재적인 위험이 있습니다. 장갑, 실험실 코트, 후드, 전신 바지, 근접 신발 등 모든 적절한 안전 관행과 개인 보호 장비를 사용해야 합니다.
1. 이중 카복실 폴리에틸렌 글리콜 (CT-PEG)33의 합성
<ol><li>1.46 g (14.6 mmol) 수치 안 수 화물 (SA) 및 209 mg (1.71 mmol) 4-디메틸 아미노피리딘 (DMAP)의 100 mL 라운드 바닥 플라스크에 추가합니다.</li>
	<li>1.1단계에서 사용되는 플라스크에 15mL의 무수스테트라하이드로푸란(THF)을 추가하고 유리 스토퍼에 맞춥니다. 플라스크를 0°C에서 30분 동안 보관하십시오.</li>
	<li>폴리에틸렌 글리콜(PEG)과 트리에틸라민(TEA)의 1.8mL(12.8mmol)을 새로운 플라스크에 4.28g(4.28mmol)을 추가한다.</li>
	<li>1.3 단계에서 사용되는 플라스크에 15mL의 무수THF를 추가하고 유리 스토퍼에 맞습니다. 질소 대기 하에서 주사기를 사용하여 1.2 단계에서 사용되는 플라스크로 천천히 용액을 전달합니다.</li>
	<li>용액을 0°C에서 2시간 동안 저은 다음 밤새 실온(RT)에서 반응을 계속합니다.</li>
	<li>회전 증발기(40°C, 0.1 MPa)를 사용하여 반응 용액을 농축하고 THF 용매를 제거한다.</li>
	<li>RT에서, 1.325 g/mL 디클로로메탄(DCM)의 15mL에서 1.6단계에서 반응 용액을 용해한 다음, 강수산물(폴리에틸렌 글리콜 디산)을 얻기 위해 15mL의 콜드 다이틸 에테르(Et2O)를 첨가한다. 필터 용지를 통해 용매를 제거합니다. 참고: 강수 단계는 3배 반복될 수 있습니다.</li>
	<li>48 시간 동안 RT에서 진공 상태에서 침전을 건조시.</li>
</ol>2. PEI-g-PEG 합합체 합성
<ol><li>305.47 단계에서 CT-PEG의 mg 1.8 과 5 mL 디메틸 설산화물 (DMSO) 플라스크에 하 고 CT-PEG가 DMSO에 완전히 용해 되도록 RT에서 저어.</li>
	<li>DMSO의 5mL에서 1-(3-디메틸아미노프로필)-3-에틸카르보디미드 염산염(EDC)의 49.46 mg을 녹인 다음, 2.1단계에서 사용되는 플라스크에 용액을 추가하고 RT에서 30분 동안 저어줍니다.</li>
	<li>DMSO의 5mL에 29.69 mg의 N-하이드록시시시니미드(NHS)를 용해하고 2.1단계에서 사용되는 플라스크에 용액을 추가한다. 3 시간 동안 RT에서 계속 저어주세요.</li>
	<li>DMSO의 10mL에 폴리에틸렌네이민(PEI)의 28.6 μL을 용해하고 2.1단계에서 사용되는 플라스크에 액액을 드롭방향으로 추가한다. 적어도 3 일 동안 저어주세요.</li>
	<li>반응형 용액을 2.4단계에서 투석 백(1,000개의 분자량 컷오프[MWCO]으로 전송합니다. 투석 가방을 투석물로 500mL초순수로 1L 비커에 넣습니다. 3 일 동안 12 시간마다 초순수를 변경합니다.</li>
	<li>다른 투석 가방 (10,000 MWCO)에 2.5 단계에서 솔루션을 전송합니다. 투석 가방을 투석물로 500mL초순수로 1L 비커에 넣습니다. 3 일 동안 12 시간마다 초순수를 변경합니다.</li>
	<li>회전 증발기(40°C, 0.1 MPa)를 이용하여 2.6단계에서 용액을 농축하고 시료를 동결건조하여 PEI-g-PEG 분말을 획득한다.</li>
</ol>3. PEI-g-PEG@AuNPs 합성
<ol><li>5 mg의 완고한 PEI-g-PEG(2.7단계)를 5mL의 초순수로 녹여 서 새로운 플라스크에 넣고 유리 스토퍼에 맞춥니다.</li>
	<li>플라스크에 0.3 mM HAuCl4 용액의 100 mL을 추가하고 RT에서 3 시간 용용액을 저어줍니다. 참고: 솔루션의 색상은 노란색에서 주황색으로 즉시 변경되어야 합니다.</li>
	<li>플라스크에 1 mg /mL NaBH4 용액의 1 mL을 추가하고 RT에서 3 시간 용용액을 저어줍니다. 참고: 반응 솔루션은 즉시 부르고뉴로 바꿔야 합니다.</li>
	<li>PEI-g-PEG@AuNPs 용액을 얻기 위해 2.5단계에서 설명된 바와 같이 3일간 투석백(1,000MWCO)을 이용하여 반응 제품을 투석한다.</li>
</ol>4. DOX-g-PEI-g-PEG@AuNPs 합성
<ol><li>2.2 mg / mL DOX 솔루션의 1 mL과 새로운 플라스크에 PEI-g PEG@AuNPs 솔루션 20 mL을 추가하고 유리 스토퍼에 맞게.</li>
	<li>초순수 1mL에 EDC 0.727 mg을 용해하고 4.1 단계에서 사용되는 플라스크에 EDC 용액을 추가한다.</li>
	<li>초순수 1mL에 NHS 0.437 mg을 용해하십시오. FLASK에 NHS 솔루션을 추가하고 1 시간 동안 RT에서 저어.</li>
	<li>2.5단계에서 설명한 대로 3일간 투석백(1,000MWCO)을 사용하여 반응 제품을 투석하여 DOX-g-PEI-g-PEG@AuNPs 솔루션을 획득하였다.</li>
</ol>5. AS1411-g-DOX-g-PEI-g-PEG@AuNPs 합성
<ol><li>새로운 플라스크에 DOX-g-PEI-g-PEG@AuNPs 용액 20mL및 AS1411(OD = 광학 밀도; 1OD ≈ 33 μg)의 4OD를 추가합니다.</li>
	<li>초순수 1mL에 28.76 mg의 EDC를 녹이고 5.1단계에서 사용되는 플라스크에 EDC 용액을 추가한다.</li>
	<li>초순수 1mL에 NHS 17.27 mg을 녹입니다. 5.1 단계에서 사용되는 플라스크에 NHS 솔루션을 추가하고 RT에서 1h에 대한 반응을 저어줍니다.</li>
	<li>AS1411-g-DOX-g-PEI-g-PEG@AuNPs 획득하기 위해 2.5단계에서 설명된 바와 같이 3일간 투석백(1,000 MWCO)을 사용하여 반응 제품을 투석한다.</li>
</ol>6. 샘플 특성화
<ol><li>핵 자기 공명(NMR) 튜브에서 클로로폼-d에서 CT-PEG 폴리머(Step 1.8) 및 PEI-g-PEG 중합체(step 2.7)를 각각 용해한다. 14.09 T 초전도 자석과 5.0mm 600 MHz 광대역 Z 그라데이션 고해상도 프로브가 장착된 600 MHz NMR 분광계를 사용하여 샘플을 분석하여 화학구조(34)를확인한다.</li>
	<li>AuNPs, DOX 및 AS1411을 분산시키고, 울트라퓨어워터에서 각각 PEI-g-PEG@AuNPs(3.4단계), DOX-g-PEI-g-PEG@AuNPs(단계 4.4), 및 AS1411-g-DOX-g-PEI-g-PEG@AuNPs(단계 5.4)를 제조하였다. 그런 다음 UV-vis 분광광계를 사용하여 큐벳으로 옮기고 자외선 가시성 (UV-vis) 스펙트럼을 기록하십시오.</li>
	<li>알루미늄 호일에 양면 접착제(~2mm x 2mm)를 부착하고 샘플 용액(PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs, AS1411 g-DOX-g-PEI-g-PEG@AuNPs)을 전체 테이프에 균일하게 찍어 보세요. X선 광전자 분광분석기를 사용하여 샘플을 분석합니다.</li>
	<li>PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs, AS1411-g-DOX-g-PEI-g-PEG@AuNPs 솔루션을 각각 초순수수에 분산시한다. 그런 다음, 큐벳으로 옮기고 동적 광 산란을 사용하여 크기 분포를 평가합니다.</li>
	<li>PEI-g-PEG@AuNPs, DOX-g-PEI-g-PEG@AuNPs, 및 AS1411 g-DOX-g-PEI-g-PEG@AuNPs 용액을 초순수 수분(각 시료당 5mL당 1방울의 샘플). 2 시간 동안 초음파 처리합니다. 구리 그리드를 샘플 솔루션에 담그고 적외선 램프 아래에서 건조시하십시오. 변속기 현미경을 사용하여 형태학을 특성화한다.</li>
	<li>AS1411-g-DOX-g-PEI-g-g-PEG@AuNPs 20kDa MWCO 투석 카세트에 1 mg을 주입한 다음, 5% 소세럼 알부민(BSA)이 있는 인산완충식살린(PBS)의 80mL에 넣습니다. 37°C에서 저어줍니다.</li>
	<li>미리 정해진 시점에서 100μL 알리쿼트를 수집하고 신선한 PBS로 대체합니다. UV-vis 분광계를 사용하여 알리쿼트의 DOX 형광 강도를 측정합니다.</li>
</ol>7. AS1411-g-DOX-g-PEI-g-PEG@AuNPs 나노 입자의 CCK-8 분석
<ol><li>Dulbecco의 수정된 독수리 매체(DMEM)에서 A549 세포를 성장시키는 것은 10% 태아 소 혈청, 100 U/mL 페니실린, 100 μg/mL 연쇄절제술로 37°C에서 95%의 공기와 5% CO2의 가습대기 하에서 보충됩니다. 배양 배지를 2일마다 교체합니다. 세포 증식 및 세포 독성 분석에 대해 5통로에서 세포를 사용하여 준비된 AS1411-g-DOX-g-PEI-g-PEG@AuNPs 나노입자의 세포 독성을 정량적으로 평가한다.</li>
	<li>각 우물에 1mL의 세포 배지를 함유하고 있는 나노입자 용액 100μL을 추가합니다. 24h 및 48h를 배양한 후, 세포 배양 판에서 배양 매체를 제거한 다음, 300 μL의 신선한 배양 배지와 30 μL의 세포 계수 키트-8(CCK-8) 키트 솔루션을 각 웰에 즉시 첨가한다. 37°C에서 CO2 인큐베이터에서 4시간 동안 배양한다.</li>
	<li>반응 솔루션 200 μL을 7.2 단계에서 96 웰 플레이트로 옮킨다. 마이크로 플레이트 판독기와 함께 570 nm에서 각 웰의 광학 밀도(OD)를 읽습니다.</li>
	<li>현미경으로 24 h 및 48 h에서 세포의 형태를 관찰하십시오.</li>
</ol>

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K., Ma, P. 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저자는 공개 할 것이 없습니다.

1H NMR스펙트럼(도 2)은CT-PEG 중합체 및 PEI-g-PEG 중합체의 성공적인 합성을 확인합니다. PEG와 PEI의 분자량은 각각 1,000및 1,200이었습니다. 또한, EDC/NHS 촉매 시스템은 아미드 반응을 통해 PEI-g-PEG 합합체를 합성하는 데 사용되었습니다. PEG와 PEI의 분자량이 PEI-g-PEG 합합체를 합성하기 위해 변경되면 반응 시간과 촉매 시스템을 재평가해야 합니다. 또한, AUNPs에 대한 PEI-g-PEG ?...

전 세계적으로 주요 공중 보건 문제이기 때문에 암은 치료율이 낮고 재발률이 높고 사망률이 높은 것으로 널리 특징지으며사망률은 1,2. 현재 종래의 항암 방법은 수술, 화학요법 및 방사선 요법3을포함하며, 그 중 화학요법은 클리닉4에서암 환자를 위한 1차 치료법이다. 임상중항암제는 주로 파클리탁셀(PTX)5 및 독소루비신(DOX)6,7을포함한다. 항신성형치료제인 DOX는 암 세포독성 및 암세포 증식 억제의 장점으로 임상화학요법에 광범위하게 적용되어왔다8,9. 그러나 DOX는 심장독성을 10,11로유발하며, DOX의 짧은 반감기는클리닉(12)에서의적용을 제한한다. 따라서, 분해가능한 약물 운반체는 DOX를 적재하고 통제된 방식으로 표적 영역으로 안전하게 방출하기 위해 필요합니다.
나노 입자는 표적 약물 전달 시스템에 널리 사용되어 왔으며 암 치료에 몇 가지 장점이 있습니다 (즉, 크기가 큰 표면 대 부피 비율, 작은 크기, 다양한 약물을 캡슐화하는 능력 및 튜닝 표면 화학 등). 13,14,15. 특히, 금 나노입자(AuNPs)는 광열암치료제(16,17)와같은 생물학적 및 생체 의학 응용 분야에서 널리 사용되고 있다. 촉진 합성 및 일반 표면 기능화와 같은 AuNPs의 독특한 특성은 암치료(18)의임상 분야에서 우수한 전망을 가지고 있다. 또한, AuNPs는 약물 전달 전략을 식별하고, 종양을 진단하고, 많은 연구에서 저항을 극복하기 위해 사용되어 왔다19,20.
하지만, AuNPs는 표적화 및 접근성 특성과 같은 향상된 투과 및 보존(EPR)을 통해 종양 병변에서 높은 국소 방출을 통해 약물 내성을 극복하기 위해 더욱 맞춤화되어야 합니다. 중합체 기능성 AuNPs는 소수성 항암제의 용용성 향상 및 장기간 순환시간(21,22)과같은 독특한 장점을 나타내었다. 폴리에틸렌 글리콜(PEG), 폴리에틸렌리민(PEI), 히알루론산, 헤파린, 크산탄껌 등 다양한 생체적합성 폴리머가 AuNP 코팅에 사용되어 왔다. 그런 다음 AuNP의 페이로드뿐만 아니라 안정성이 잘향상됩니다(23). 구체적으로, PEI는 1차, 이차 및 고등아민(24)의많은 반복 단위로 구성된 고도로 분진된 중합체이다. PEI는 우수한 용해도, 낮은 점도 및 높은 수준의 기능을 가지고 있으며, 이는 AuNPs코팅에 적합합니다.
한편, 항암제는 하중 효율이 향상되어 암세포에 직접 전달되어야 하며, 1차 및 첨단 전이성 종양 을 치료하기 위한 독성이 낮을 수있다(25). 표적 리간드항은 항암제 표적 전달시스템(26)에큰 잠재력을 가지고 있다. 표적 분자 결합에 대한 선택성은 특이성을 표적으로 하는 항암제를 부여하고 병성 조직에 있는 약 농축을 증가합니다(27). 더 많은 리간드는 항체, 폴리펩티드 및 소분자를 포함한다. 다른 리간드에 비해, 핵산 aptamers는 체외에서 합성될 수 있고 수정하기 쉽습니다. AS1411은 암세포28,29,30에과발현 표적 핵단백질 수용체에 구체적으로 결합하기 위해 안정된 조광 G-테트라머 구조를 형성하는 수정되지 않은 26bp 인포디스터 올리고뉴클레오티드이다. AS1411은 많은 암세포의 증식을 억제하지만 건강한세포(31,32)의성장에영향을 미치지 않는다. 그 결과, AS1411은 이상적인 표적 약물 전달 시스템을 제조하는 데 사용되어 왔다.
본 연구에서는, PEI-g-PEG 공중합체는 아마이드 반응을 통해 합성되고, PEI-g-PEG 공합체 코팅된 금 나노입자(PEI-g-PEG@AuNPs)가 제조된다. 또한 DOX 및 AS1411은 도 1에도시된 바와 같이 준비된 PEI-g-PEG@AuNPs 순차적으로 연결된다. 이 상세한 프로토콜은 연구원이 DOX 및 AS1411로로드된 새로운 PEI-g-PEG@AuNPs 제조와 관련된 많은 일반적인 함정을 피할 수 있도록 돕기 위한 것입니다.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>4-Dimethylaminopyridine</td>
			<td>Macklin</td>
			<td>D807273</td>
			<td></td>
		</tr>
		<tr>
			<td>A549 cell</td>
			<td></td>
			<td>ATCC CCL-185TM</td>
			<td></td>
		</tr>
		<tr>
			<td>AS1411</td>
			<td>BBI Life Sciences Corporation</td>
			<td></td>
			<td>5&#39;-d (TTTGGTGGTGGTGGTTGTGGTGGTGGTGG) FL-AS1411 (fluorophore-labeled AS1411)</td>
		</tr>
		<tr>
			<td>Anhydrous Tetrahydrofuran (THF)</td>
			<td>SinoPharm Chemical Reagent Co., Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cell counting kit-8 (CCK-8)</td>
			<td>Sigma Aldrich</td>
			<td>96992-500TESTS-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichloromethane</td>
			<td>Traditional Chinese medicine</td>
			<td>80047318</td>
			<td></td>
		</tr>
		<tr>
			<td>Diethyl ether (Et2O)</td>
			<td>SinoPharm Chemical Reagent Co., Ltd</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dimethyl sulfoxide</td>
			<td>Macklin</td>
			<td>D806645</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified Eagle&#39;s medium (DMEM)</td>
			<td>Sigma Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Doxorubicin hydrochloride</td>
			<td>Rhawn</td>
			<td>R017518</td>
			<td></td>
		</tr>
		<tr>
			<td>Ether absolute</td>
			<td>Traditional Chinese medicine</td>
			<td>80059618</td>
			<td></td>
		</tr>
		<tr>
			<td>Field Emission Transmission Electron Microscope</td>
			<td>FEI Company</td>
			<td>Tecnai G2 F 20</td>
			<td></td>
		</tr>
		<tr>
			<td>Gold(III) chloride trihydrate</td>
			<td>Rhawn</td>
			<td>R016035</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Particle-size Instrument</td>
			<td>Malvern Instruments Ltd</td>
			<td>ZetasizerNanoZS/Masterszer3000E</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate Reader</td>
			<td>Molecular Devices</td>
			<td>SpectraMax 190</td>
			<td></td>
		</tr>
		<tr>
			<td>N-(3-Dimethylaminopropyl)-N&#39;-ethylcarbodiimide hydrochloride</td>
			<td>Macklin</td>
			<td>N808856</td>
			<td></td>
		</tr>
		<tr>
			<td>N-Hydroxysuccinimide</td>
			<td>Macklin</td>
			<td>H6231</td>
			<td></td>
		</tr>
		<tr>
			<td>NMR software</td>
			<td>Delta 5.2.1</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nuclear Magnetic Resonance Spectrometer</td>
			<td>JEOL</td>
			<td>JNM-ECZ600R/S3</td>
			<td></td>
		</tr>
		<tr>
			<td>Origin 8.5</td>
			<td>OriginLab</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin</td>
			<td>Sigma Aldrich</td>
			<td>V900929-100ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate-buffered saline</td>
			<td>Sigma Aldrich</td>
			<td>P4417-100TAB</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly(ethylene glycol)</td>
			<td>Sigma Aldrich</td>
			<td>81188</td>
			<td>BioUltra, average Mn ~ 1000</td>
		</tr>
		<tr>
			<td>Poly (ethyleneimine) solution</td>
			<td>Sigma Aldrich</td>
			<td>482595</td>
			<td>average Mn ~ 1200, 50 wt.% in H2O</td>
		</tr>
		<tr>
			<td>Sodium borohydride, powder</td>
			<td>Acros</td>
			<td>C18930</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptomycin</td>
			<td>Sigma Aldrich</td>
			<td>85886-10ML</td>
			<td></td>
		</tr>
		<tr>
			<td>Succinic anhydride</td>
			<td>Traditional Chinese medicine</td>
			<td>30171826</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetrahydrofuran</td>
			<td>Traditional Chinese medicine</td>
			<td>40058161</td>
			<td></td>
		</tr>
		<tr>
			<td>Triethylamine</td>
			<td>Traditional Chinese medicine</td>
			<td>80134318</td>
			<td></td>
		</tr>
		<tr>
			<td>UV/VIS/NIR Spectrometer</td>
			<td>Lambda950</td>
			<td>Lambda950</td>
			<td></td>
		</tr>
		<tr>
			<td>X-ray Photoelectron Spectrometer</td>
			<td>Thermo&#8194;Fisher&#8194;Scientific</td>
			<td>K-ALPHA 0.5EV</td>
			<td></td>
		</tr>
	</tbody>
</table>

synthesis of aptamer-pei-g-peg modified gold nanoparticles loaded with doxorubicin for targeted drug delivery

건강한 세포에서 약물 내성 및 독성으로 인해 독소루비신(DOX)의 사용은 임상 암 요법에서 제한적입니다. 이 프로토콜은 아미드 반응을 통해 로드된 aptamer(AS1411) 및 DOX를 탑재한 폴리에틸렌 글리콜(PEI-g-PEG) 중합체 기능성 금 나노입자(AuNPs)로 이식된 폴리(에틸레니민)의 설계를 설명한다. AS1411은 특히 DOX가 건강한 세포 대신 암세포를 표적으로 할 수 있도록 암세포에 표적 뉴클레놀린 수용체와 결합된다. 첫째, PEG는 카박스화되어 1HNMR 분석에 의해 확인된 PEI-g-PEG 중합체를 얻기 위해 분기된 PEI에 이식된다. 다음으로, PEI-g-PEG 공합체 코팅 금 나노입자(PEI-g-PEG@AuNPs)가 합성되고, DOX 및 AS1411은 아마드 반응을 통해 점차적으로 AuNPs에 공유결합된다. 준비된 AS1411-g-DOX-g-PEI-g-PEG@AuNPs 의 직경은 ~39.9nm이며, 제타 전위 -29.3mV로, 나노입자가 물과 세포 배지에서 안정되어 있음을 나타낸다. 세포 세포 독성 연구는 새로 설계된 DOX로드 AuNPs가 암세포 (A549)를 죽일 수 있음을 보여줍니다. 이 합성은 순차적 인 아미데 반응에 의해 달성되는 AuNPs에 PEI-g-PEG 공중 합합체, aptamers 및 DOX의 섬세한 배열을 보여줍니다. 이러한 aptamer-PEI-g-PEG 기능화된 AuNPs는 암 치료에서 표적 약물 전달을 위한 유망한 플랫폼을 제공합니다.

1 H NMR 분광법은 CT-PEG 중합체 및 PEI-g-PEG 중합체(도2)의성공적인 합성을 확인하기 위해 사용되었다. 도 2a는 δ = 3.61 ppm 및 carboxyl 양성자 신호에서 메틸렌 양성자 신호가 δ = 2.57 ppm에서 CT-PEG 폴리머의 성공적인 합성을 확인함을 나타낸다. 도 2b는 δ = 2.6 ppm 및 peI의 양성자 신호에서 PEG의 메틸렌 양?...

Watch this Scientific Journal Video about Synthesis of Aptamer-PEI-g-PEG Modified Gold Nanoparticles Loaded with Doxorubicin for Targeted Drug Delivery at JoVE.com

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

이 프로토콜에서, 독소루비신로드 AS1411-g-PEI-g-PEG 변형 금 나노 입자는 3 단계 아마이드 반응을 통해 합성된다. 이어서, 독소루비신은 암 치료를 위한 암세포를 표적으로 하기 위하여 장전되고 전달된다.

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 ...

synthesis-aptamer-pei-g-peg-modified-gold-nanoparticles-loaded-with

Research

JoVE Journal

Chemistry

5.8K 조회수.  Xinyang Normal University. 이 프로토콜에서, 독소루비신로드 AS1411-g-PEI-g-PEG 변형 금 나노 입자는 3 단계 아마이드 반응을 통해 합성된다. 이어서, 독소루비신은 암 치료를 위한 암세포를 표적으로 하기 위하여 장전되고 전달된다.

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

Template Directed Synthesis of Plasmonic Gold Nanotubes with Tunable IR Absorbance

A nearly parallel array of pores can be produced by anodizing aluminum foils in acidic environments1, 2. Applications of anodic aluminum oxide (AAO) membranes have been under development since the 1990's and have become a common method to template the synthesis of high aspect ratio nanostructures, mostly by electrochemical growth or pore-wetting. Recently, these membranes have become commercially available in a wide range of pore sizes and densities, leading to an extensive library of functional nanostructures being synthesized from AAO membranes. These include composite nanorods, nanowires and nanotubes made of metals, inorganic materials or polymers 3-10. Nanoporous membranes have been used to synthesize nanoparticle and nanotube arrays that perform well as refractive index sensors, plasmonic biosensors, or surface enhanced Raman spectroscopy (SERS) substrates 11-16, as well as a wide range of other fields such as photo-thermal heating 17, permselective transport 18, 19, catalysis 20, microfluidics 21, and electrochemical sensing 22, 23. Here, we report a novel procedure to prepare gold nanotubes in AAO membranes. Hollow nanostructures have potential application in plasmonic and SERS sensing, and we anticipate these gold nanotubes will allow for high sensitivity and strong plasmon signals, arising from decreased material dampening 15.

Solution-suspendable gold nanotubes with controlled dimensions can be synthesized by electrochemical deposition in porous anodic aluminum oxide (AAO) membranes using a hydrophobic polymer core. Gold nanotubes and nanotube arrays hold promise for applications in plasmonic biosensing, surface-enhanced Raman spectroscopy, photo-thermal heating, ionic and molecular transport, microfluidics, catalysis and electrochemical sensing.

템플릿 조정할 IR의 흡광도와 Plasmonic 골드 나노 튜브의 합성을 감독

A nearly parallel array of pores can be produced by anodizing aluminum foils in acidic environments1, 2. Applications of anodic aluminum oxide (AAO) ...

연구

화학

Preparation and Characterization of SDF-1&#945;-Chitosan-Dextran Sulfate Nanoparticles

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under appropriate conditions. The glycan nanoparticles are useful carriers for protein factors, which facilitate the in vivo delivery of the proteins and sustain their retention in the targeted tissue. The glycan polyelectrolyte complexes are also ideal for protein delivery, as the incorporation is carried out in aqueous solution, which reduces the likelihood of inactivation of the proteins. Proteins with a heparin-binding site adhere to dextran sulfate readily, and are, in turn, stabilized by the binding. These particles are also less inflammatory and toxic when delivered in vivo. In the protocol described below, SDF-1&#945; (Stromal cell-derived factor-1&#945;), a stem cell homing factor, is first mixed and incubated with dextran sulfate. Chitosan is added to the mixture to form polyelectrolyte complexes, followed by zinc sulfate to stabilize the complexes with zinc bridges. The resultant SDF-1&#945;-DS-CS particles are measured for size (diameter) and surface charge (zeta potential). The amount of the incorporated SDF-1&#945; is determined, followed by measurements of its in vitro release rate and its chemotactic activity in a particle-bound form.

Preparation and Characterization of SDF-1&amp;#945;-Chitosan-Dextran Sulfate Nanoparticles

The objective of this protocol is to incorporate SDF-1&#945;, a stem cell homing factor, into dextran sulfate-chitosan nanoparticles. The resultant particles are measured for their size and zeta potential, as well as the content, activity, and in vitro release rate of SDF-1&#945; from the nanoparticles.

SDF-1α-키토산 덱스 트란 황산 나노 입자의 제조 및 특성

Chitosan (CS) and dextran sulfate (DS) are charged polysaccharides (glycans), which form polyelectrolyte complex-based nanoparticles when mixed under ...

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 ...

Hydroquinone Based Synthesis of Gold Nanorods

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the synthetic methods for the preparation of gold nanorods are still not fully optimized. In this paper we describe a new, highly efficient, two-step protocol based on the use of hydroquinone as a mild reducing agent. Our approach allows the preparation of nanorods with a good control of size and aspect ratio (AR) simply by varying the amount of hexadecyl trimethylammonium bromide (CTAB) and silver ions (Ag+) present in the "growth solution". By using this method, it is possible to markedly reduce the amount of CTAB, an expensive and cytotoxic reagent, necessary to obtain the elongated shape. Gold nanorods with an aspect ratio of about 3 can be obtained in the presence of just 50 mM of CTAB (versus 100 mM used in the standard protocol based on the use of ascorbic acid), while shorter gold nanorods are obtained using a concentration as low as 10 mM.

This paper describes a protocol for the synthesis of gold nanorods, based on the use of hydroquinone as reducing agent, plus the different mechanisms for controlling their size and aspect ratio.

금 나노 막대의 하이드로 퀴논 기반 합성

Gold nanorods are an important kind of nanoparticles characterized by peculiar plasmonic properties. Despite their widespread use in nanotechnology, the ...

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

We demonstrate a method for the synthesis of NixNb1-xO catalysts with sponge-like and fold-like nanostructures. By varying the Nb:Ni ratio, a series of NixNb1-xO nanoparticles with different atomic compositions (x = 0.03, 0.08, 0.15, and 0.20) have been prepared by chemical precipitation. These NixNb1-xO catalysts are characterized by X-ray diffraction, X-ray photoelectron spectroscopy, and scanning electron microscopy. The study revealed the sponge-like and fold-like appearance of Ni0.97Nb0.03O and Ni0.92Nb0.08O on the NiO surface, and the larger surface area of these NixNb1-xO catalysts, compared with the bulk NiO. Maximum surface area of 173 m2/g can be obtained for Ni0.92Nb0.08O catalysts. In addition, the catalytic hydroconversion of lignin-derived compounds using the synthesized Ni0.92Nb0.08O catalysts have been investigated.

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

A protocol for the synthesis of sponge-like and fold-like Ni1-xNbxO nanoparticles by chemical precipitation is presented.

Nb2O5 의 합성을 위한 화학 강 수 방법을 수정할 높은 특정 표면 영역으로 대량 니켈 촉매

We demonstrate a method for the synthesis of NixNb1-xO catalysts with sponge-like and fold-like nanostructures. By varying the Nb:Ni ratio, a series of ...

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 ...

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery

Reversible addition-fragmentation chain transfer (RAFT) polymerization integrates the advantages of radical polymerization and living polymerization. This work presents the preparation of methionine functionalized biocompatible block copolymers via RAFT polymerization. Firstly, N,N-bis(2-hydroxyethyl)methacrylamide-b-N-(3-aminopropyl)methacrylamide (BNHEMA-b-APMA, BA) was synthesized via RAFT polymerization using 4,4&#8217;-azobis(4-cyanovaleric acid) (ACVA) as an initiating agent and 4-cyanopentanoic acid dithiobenzoate (CTP) as the chain transfer agent. Subsequently, N,N-bis(2-hydroxyethyl)methacrylamide-b-N-(3-guanidinopropyl)methacrylamide (methionine grafted BNHEMA-b-GPMA, mBG) was prepared by modifying amine groups in APMA with methionine and guanidine groups. Three kinds of block polymers, mBG1, mBG2, and mBG3, were synthesized for comparison. A ninhydrin reaction was used to quantify the APMA content; mBG1, mBG2, and mBG3 had 21%, 37%, and 52% of APMA, respectively. Gel permeation chromatography (GPC) results showed that BA copolymers possess molecular weights of 16,200 (BA1), 20,900(BA2), and 27,200(BA3) g/mol. The plasmid DNA (pDNA) complexing ability of the obtained block copolymer gene carriers was also investigated. The charge ratios (N/P) were 8, 16, and 4 when pDNA was complexed completely with mBG1, mBG2, mBG3, respectively. When the N/P ratio of mBG/pDNA polyplexes was higher than 1, the Zeta potential of mBG was positive. At an N/P ratio between 16 and 32, the average particle size of mBG/pDNA polyplexes was between 100-200 nm. Overall, this work illustrates a simple and convenient protocol for the block copolymer carrier synthesis.

This work presents the preparation of methionine functionalized biocompatible block copolymers (mBG) via the reversible addition-fragmentation chain transfer (RAFT) method. The plasmid DNA complexing ability of the obtained mBG and their transfection efficiency were also investigated. The RAFT method is very beneficial for polymerizing monomers containing special functional groups.

표적 플라스미드 DNA 전달을 위한 메티오닌 기능화 생체 적합성 블록 공중합체

Reversible addition-fragmentation chain transfer (RAFT) polymerization integrates the advantages of radical polymerization and living polymerization. This ...

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 ...

Gold Nanoparticle Modified Carbon Fiber Microelectrodes for Enhanced Neurochemical Detection

For over 30 years, carbon-fiber microelectrodes (CFMEs) have been the standard for neurotransmitter detection. Generally, carbon fibers are aspirated into glass capillaries, pulled to a fine taper, and then sealed using an epoxy to create electrode materials that are used for fast scan cyclic voltammetry testing. The use of bare CFMEs has several limitations, though. First and foremost, the carbon fiber contains mostly basal plane carbon, which has a relatively low surface area and yields lower sensitivities than other nanomaterials. Furthermore, the graphitic carbon is limited by its temporal resolution, and its relatively low conductivity. Lastly, neurochemicals and macromolecules have been known to foul at the surface of carbon electrodes where they form non-conductive polymers that block further neurotransmitter adsorption. For this study, we modify CFMEs with gold nanoparticles to enhance neurochemical testing with fast scan cyclic voltammetry. Au3+ was electrodeposited or dipcoated from a colloidal solution onto the surface of CFMEs. Since gold is a stable and relatively inert metal, it is an ideal electrode material for analytical measurements of neurochemicals. Gold nanoparticle modified (AuNP-CFMEs) had a stability to dopamine response for over 4 h. Moreover, AuNP-CFMEs exhibit an increased sensitivity (higher peak oxidative current of the cyclic voltammograms) and faster electron transfer kinetics (lower &#916;EP or peak separation) than bare unmodified CFMEs. The development of AuNP-CFMEs provides the creation of novel electrochemical sensors for detecting fast changes in dopamine concentration and other neurochemicals at lower limits of detection. This work has vast applications for the enhancement of neurochemical measurements. The generation of gold nanoparticle modified CFMEs will be vitally important for the development of novel electrode sensors to detect neurotransmitters in vivo in rodent and other models to study neurochemical effects of drug abuse, depression, stroke, ischemia, and other behavioral and disease states.

In this study, we modify carbon-fiber microelectrodes with gold nanoparticles to enhance the sensitivity of neurotransmitter detection.

향상된 신경 화학적 검출을 위한 금 나노입자 수정 탄소 섬유 마이크로 전극

For over 30 years, carbon-fiber microelectrodes (CFMEs) have been the standard for neurotransmitter detection. Generally, carbon fibers are aspirated into ...

Gold Nanoparticle Synthesis

Gold nanoparticles (Au nanoparticles) that are ~12 nm in diameter were synthesized by rapidly injecting a solution of 150 mg (0.15 mmol) of tetrachloroauric acid in 3.0 g (3.7 mmol, 3.6 mL) of oleylamine (technical grade) and 3.0 mL of toluene into a boiling solution of 5.1 g (6.4 mmol, 8.7 mL) of oleylamine in 147 mL of toluene. While boiling and mixing the reaction solution for 2 hours, the color of the reaction mixture changed from clear, to light yellow, to light pink, and then slowly to dark red. The heat was then turned off, and the solution was allowed to gradually cool down to room temperature for 1 hour. The gold nanoparticles were then collected and separated from the solution using a centrifuge and washed three times; by vortexing and dispersing the gold nanoparticles in 10 mL portions of toluene, and then precipitating the gold nanoparticles by adding 40 mL portions of methanol and spinning them in a centrifuge. The solution was then decanted to remove any remaining byproducts and unreacted starting materials. Drying the gold nanoparticles in a vacuum environment produced a solid black pellet; which could be stored for long periods of time (up to one year) for later use, and then redissolved in organic solvents such as toluene.

A protocol for synthesizing ~12 nm diameter gold nanoparticles (Au nanoparticles) in an organic solvent is presented. The gold nanoparticles are capped with oleylamine ligands to prevent agglomeration. The gold nanoparticles are soluble in organic solvents such as toluene.