우리는 자신의 종류의 지원에 교수 JP 파츠 (생물 물리 화학과, 하이델베르크 대학교 및 새로운 재료 및 바이오 시스템학과, 지능형 시스템을위한 맥스 플랑크 연구소, 슈투트가르트) 감사합니다. 막스 플랑크 먼과 도이치 Forschungsgemeinschaft (EAC-A에 DFG SFB/TR79.)에서 재정 지원도 크게 인정합니다.

1. 11-Mercaptoundecanoyl-N-히드 록시 에스테르 (MU-NHS)의 합성 <ol><li> 실온 (RT)에서 40 ㎖ 디클로로 메탄 (PA)에있는 10 ㎖의 아세톤 (PA)에있는 (디메틸 아미노) 피리딘 1g에 11 mercaptoundecanoic 산 - 500 mg의 N-하이드 록시의 솔루션과 30 mg의 4 적하 추가합니다. </li><li> 0 ° C에 대한 반응을 냉각과 적하 1.1 g의 N (질소 분위기 하에서) 10 ㎖ 용액에 N &#39;- 디시 클로 헥실를 추가합니다. 1 시간 동안 저온에서 반응을 계속 한 후 실온에서 밤새 교반 하였다. </li><li> 침전물을 여과하고 감압 하에서 건조. 1:1 비율 석유 벤젠, 에틸 아세테이트 (PA)으로 플래쉬 크로마토 그래피에 의해 생성물을 정제 하였다. </li></ol> 2. 동종 골드 레이어의 준비 <ol><li> 정밀 물티슈 및 에틸 아세테이트 (PA) 및 메탄올 (PA)의 1:1 혼합물을 함유하는 용액에 5 분 동안 초음파 처리들을 깨끗한 유리 커버 슬립. RINS전자 메탄올 기판 및 질소 흐름 하에서 건조. </li><li> 스퍼터 코팅 장치의 깨끗한 기판을 놓습니다. 챔버 대피. 60 초간 스퍼터링에 의해 크롬 타겟으로부터 최상부의 크롬 산화물 층을 제거한다. 금속 빔과 40 nm의 금 층으로 코팅 한 다음에 10 nm의 크롬 층과 코트를 아래의 샘플을 놓습니다. </li></ol> 3. BMP-2의 표면 고정화 <ol><li> 약 1 ㎜의 최종 농도에서 N MU-NHS, N-dimethlyformamide (DMF)에 용해시키고, 질소 대기 하에서 4 시간 동안 RT에서 MU-NHS 용액에 금 기판을 부화. </li><li> 2 분 DMF에 초음파 처리 표면은, DMF 및 MeOH로 세척하고 질소 흐름 하에서 건조. </li><li> 100 ㎍ / ㎖ (-80 ° C에서 보관 분량 씩)의 농도로 멸균 4 mM의 염산에 rhBMP-2의 주식 솔루션을 준비합니다. 작업 희석액 (3.5 ㎍ / ㎖)의 경우, PBS / 염화나트륨 (PBS 1 M NaCl을 함유)과의 원액을 희석사용하기 직전에 pH를 8.5로 djust. </li><li> 4 ° C 하룻밤에 rhBMP-2 작업 솔루션에 MU-NHS를 기능화 표면을 품어. 배양 상층 액을 제거합니다. 초음파 처리 2 분 동안 PBS / 염화나트륨의 표면과 멸균 PBS / NaCl로 3 배 씻는다. </li></ol> 4. 표면 특성 <ol><li> 항체의 비특이적 흡착을 차단하기 위해, 5 % BSA와 표면을 품어 (w / v)의 안티 - BMP-2 항체 (1:100 1 % (W / V를 배양 한 다음 실온에서 1 시간을위한 PBS 용액 RT에서 1 시간 동안) BSA / PBS 용액). PBS와 30 초 동안 초음파 처리로 두 번 표면을 세척 할 것. </li><li> 다음 PBS로 두 번 씻어 RT에서 30 분 동안 HRP - 복합 이차 항체 (1:1,000 1 % (W / V) BSA / PBS 용액)으로 기판을 품어. </li><li> 플레이트 리더로 ​​570 nm에서 HRP의 효소 활성을 측정하기 위해 Ampliflu 레드 분석을 사용합니다. </li></ol> 5. 생물학적 활성 분석 <ol><li> 6 자 PL 잘 당 종자 1 × 10 5 마우스 C2C12의 근육 아세포10 % FBS, 1 % 페니실린 / 스트렙토 마이신 및 37 ° C / 5 % CO 2에서 24 시간 동안 배양들을 보충 된 피루브산을 함유하는 하이 글루코스 DMEM 이루어진 증식 배지에서 ATES. </li><li> 고정 된 BMP-2 장식 표면에 파종하기 전에 3-5 시간 동안 혈청이없는 DMEM 세포를 굶어. </li><li> 단기 신호 유도의 조사를 위해, 200 ㎕의 신선한 혈청이없는 DMEM에 의해 매체를 교체하고 세포 위의 고정 된 BMP-2 표면을 배치합니다. 표면이 긁히지 않도록 좋은 팁 핀셋으로 취급되어야한다. </li><li> 부드럽게 표면을 제거하고 피펫 매체를 대기음, PBS로 두 번 세포를 씻으십시오. 세포 반응의 분석을 진행합니다. </li><li> 장기 생물학적 활성, 플레이트 세포의 분석 상 6 일 동안 낮은 혈청 조건 (2 % FBS)에서 37 ° C / 5 % CO 2의 표면과 문화를 그들. </li></ol>

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Bioeng. 98 (6), 1109-1122 (2007).</li><li>Puleo, D., Kissling, R., Sheu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+to+immobilize+bioactive+proteins,+including+bone+morphogenetic+protein-4+(BMP-4),+on+titanium+alloy.">A technique to immobilize bioactive proteins, including bone morphogenetic protein-4 (BMP-4), on titanium alloy.</a> Biomaterials. 23 (9), 2079-2087 (2002).</li></ol>

저자가 공개하는 게 없다.

이 프로토콜에서 우리는 생리 활성 rhBMP-2와 기능화 표면의 준비에 대해 설명합니다. ; rhBMP-2 단백질의 2) 공유 고정화 금 표면에 관능 링커의 자기 조립 단분자막 (SAM)의 1) 초기 형성 :이 방법은 두 단계를 포함한다. 이전의 연구에서, 우리는 관능 링커 및 성장 인자의 효과적인 결합을 검증하고, 표면 고정화 rhBMP-2의 생물학적 활성 (19)를 유지 것을 보여 주었다. 고정화 된 형태로 제공 성장 ...

뼈 형태 형성 단백질 2 (BMP-2) 형질 전환 성장 인자 (TGF-β) 가족과 드 노보 골 형성 유도뿐만 아니라 배아 발달과 성인 동안 여러 조직의 조절기 역할을 1-3 항상성의 구성원입니다. 생물학적 활성 homodimeric BMP-2 단백질의 각 단량체는 매우 모든 BMP에 4에 보존되어있는 &quot;시스테인 매듭&quot;모티브가 포함되어 있습니다. 일곱 번째 시스테인이 두 단량체 5,6 사이의 분자간 결합을 형성, 이량에 관여하는 반면, 일곱 시스테인 잔기의 여섯, 각각의 단량체를 안정 분자 내 이황화 결합을 형성한다. 이것은 고도로 보존 된 시스테인 매듭은 BMP-2 단백질의 입체 구조를 정의하고 열, 변성제 및 산성 pH 7-9 대한 저항으로서 독특한 특성을 결정한다. BMP-2함으로써 신호 전달을 유도 세린 / 트레오닌 키나아제 막 관통 수용체 바인딩 <sup&gt; 10-12. 수용체 올리고머의 모드에 따라 서로 다른 신호 전달 경로가 활성화되어 TGF 계통에 의존하는 통로가 Smad의 복잡한 핵 전좌와의 전사 활성에있는 수용체 인산화의 결과에 의해 활성화 된 반면 Smad의 독립적 인 신호 폭포는, P38 신호를 통해 알칼리 포스 파타 아제의 유도로 연결 이러한 분화의 억제 (ID) 12 ~ 14과 같은 특정 표적 유전자. 뼈, BMP-2, 따라서 치료 및 뼈의 드 노보 형성을 자극하는 골아 세포에 중간 엽 줄기 세포의 분화를 유도한다. 현재 재조합 BMP-2가 골절 사이트의 치유를 강화하기 위하여 임상 적으로 적용되는 표현. 뼈 조직 공학의 일반적인 전략은 로컬 배달 시스템에 비해 덜 침습적 주사 성장 인자의 사용이다. 그러나, 생체 내 연구 및 임상 응용 프로그램을 보여 그 짧은 생물학적 반감기, 불특정 LOCalization와 BMP-2의 빠른 현지 통관 여러 지역 자궁외 및 조직 문제 (15)가 발생할 수 있습니다. 따라서, 효과적인 프리젠 테이션, 물질 내에서 또는 상 BMP-2의 함정 수사 또는 고정을 얻으려면 대상 사이트에서 로컬 및 지속적인 전달을 위해 필요합니다. 지속 전달은 물리적 포착, 흡착 또는 이온 착물 (16)과 같은 비공유 결합 고정 방식으로 달성 될 수있다. 그러나, 그것은 알려져있다 분자 (17)의 변성의 표면에 단백질의 비특이적 흡착 할 수있다 결과. 성장 인자의 결합을 공유 결합의 경우, 지지체의 종류가 지난 십 년간 개발되어왔다. 예를 들어 단백질의 아미노 또는 카르복시기를 대상으로 관능 링킹 분자의 사용은 반드시 그 고정화를 달성하는 단백질 수정을 요구하지 않는 방식의 한 유형이다. 사실, 반면 단백질 변형 단백질 배향을 제어 이점을 제공한다인공 도메인, 펩타이드 태그 및 부위 특이 적 쇄의 도입은 성장의 생물학적 활성은 17 요인 변경할 수있다. 따라서, 인해 담체와의 상호 작용에 변성을 회피하는, 표면은 원하는 요소 (18)의 결합에 의해 다음에 링킹 분자의 자기 조립 단분자막 (SAM), 예를 들어, 미리 관능 화 될 수있다. 우리는 그것의 유리 아민의 잔류를 대상으로 공유 표면에 BMP-2를 고정시키기 SAM-기반 접근 방식을 사용하고 고정 된 단백질의 단기 및 장기 생물학적 활성 19 두를 유지하는 것으로 나타났습니다. 이 프로토콜은 세포막에서 발생하고 골 형성 신호에 대한 책임 세포 내 신호를 조절하는 메커니즘에 체외 연구를위한 세포에 BMP-2를 제공하는 간단하고 효율적인 방법을 제공합니다.

<table border="1">
	<tbody>
		<tr>
			<td>Name of Material/ Equipment</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments/Description</td>
		</tr>
		<tr>
			<td>N-hydroxysuccinimide</td>
			<td>Sigma-Aldrich</td>
			<td>130672</td>
			<td></td>
		</tr>
		<tr>
			<td>4-(dimethylamino)pyridin</td>
			<td>Sigma-Aldrich</td>
			<td>522805</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>AppliChem</td>
			<td>A2282</td>
			<td></td>
		</tr>
		<tr>
			<td>11-mercaptoundecanoic acid</td>
			<td>Sigma-Aldrich</td>
			<td>674427</td>
			<td></td>
		</tr>
		<tr>
			<td>Dichlormethane</td>
			<td>Merck</td>
			<td>106050</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N&#39;-dicyclohexylcarbodiimide</td>
			<td>Sigma-Aldrich</td>
			<td>D80002</td>
			<td></td>
		</tr>
		<tr>
			<td>Petroleum benzene</td>
			<td>Merck</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glass coverslips</td>
			<td>Carl Roth</td>
			<td>M 875</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylacetate</td>
			<td>AppliChem</td>
			<td>A3550</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Carl Roth</td>
			<td>4627</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N-dimethylformamide</td>
			<td>Carl Roth</td>
			<td>T921</td>
			<td></td>
		</tr>
		<tr>
			<td>rhBMP-2</td>
			<td>R&#38;D Systems</td>
			<td>355-BM</td>
			<td>Carrier-free; expressed in E.coli</td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>PAA</td>
			<td>H15-002</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Carl Roth</td>
			<td>HN00.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly(dimethyl siloxane) (PDMS)</td>
			<td>Dow Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sylgard 184 silicone elastomer kit</td>
			<td>Dow Corning</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-rhBMP-2</td>
			<td>Sigma</td>
			<td>B9553</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-mouse IgG-HRP</td>
			<td>Santa Cruz</td>
			<td>sc-2005</td>
			<td>Secondary antibody</td>
		</tr>
		<tr>
			<td>Ampliflu Red assay</td>
			<td>Sigma</td>
			<td>90101</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Modified Eagle Medium (DMEM) (1x), liquid</td>
			<td>Gibco</td>
			<td>41966</td>
			<td>High glucose</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum (FBS)</td>
			<td>Sigma</td>
			<td>F7524</td>
			<td>Sterile filtered, cell culture tested</td>
		</tr>
		<tr>
			<td>Pen/Strep</td>
			<td>Gibco</td>
			<td>15140</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin 0.05% (1x) with EDTA 4Na</td>
			<td>Gibco</td>
			<td>25300</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine (0.1 M)</td>
			<td>Riedel-de Ha&#235;n</td>
			<td>33226</td>
			<td></td>
		</tr>
		<tr>
			<td>IGEPAL CA-630 (1%)</td>
			<td>Sigma</td>
			<td>I8896</td>
			<td>Lysis buffer (ALP assay)19</td>
		</tr>
		<tr>
			<td>Magnesium chloride (MgCl2)(1 mM)</td>
			<td>Carl Roth</td>
			<td>HNO3.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Zinc chloride (ZnCl2) (1 mM)</td>
			<td>Carl Roth</td>
			<td>3533.1</td>
			<td></td>
		</tr>
		<tr>
			<td>p-nitrophenylphosphate (pNPP)</td>
			<td>Sigma</td>
			<td>S0942</td>
			<td>Phosphatase substrate</td>
		</tr>
		<tr>
			<td>Anti-mysin heavy chain (MHC)</td>
			<td>Developmental Studies Hybridoma Bank, University of Iowa</td>
			<td>MF20</td>
			<td>Monoclonal antibody</td>
		</tr>
		<tr>
			<td>Alexa Fluor 488 Goat anti-mouse IgG</td>
			<td>Invitrogen</td>
			<td>A11001</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sigma</td>
			<td>D9542</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		</tr>
		<tr>
			<td>Ultrsonic bath (Sonorex Super RK 102H), Frequency 35 kHz</td>
			<td>BANDELIN electronic GmbH &#38; Co. KG</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MED 020 Sputtercoating system</td>
			<td>BAL-TEC AG</td>
			<td></td>
			<td>Coating conditions 
			Cr: 120 mA, 1.3 x 10-2 mbar, 30 sec 
			Au: 60 mA, 5.0 x 10-2 mbar, 45 sec</td>
		</tr>
		<tr>
			<td>Tecan Infinite M200 Plate reader</td>
			<td>Tecan</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

covalent binding of bmp-2 on surfaces using a self-assembled monolayer approach

뼈 형태 형성 단백질 2 (BMP-2) 뼈 조직의 세포 외 매트릭스에 포함 된 성장 인자이다. 따라서 치유와 드 노보 골 형성을 자극하는 조골 세포에 중간 엽 세포 분화의 트리거로 BMP-2의 역할. 비계와 함께 재조합 인간 BMP-2 (rhBMP-2)의 임상 프레 젠 테이션의 형태에 따라, 최근의 논쟁을 제기하고 양이 전달 될 수 있습니다. 여기에 제시된 프로토콜은 세포에 대한 생체 외 연구에 BMP-2를 제공하는 간단하고 효율적인 방법을 제공합니다. 우리는 이종 링커 이루어진 자기 조립 단분자막을 형성하는 방법을 설명하고, rhBMP-2의 공유 고정화를 구하는 후속 결합 단계를 나타낸다. 이 방법으로는 단백질의 생물학적 활성을 유지하면서, BMP-2의 지속적인 프레젠테이션을 달성하는 것이 가능하다. 사실, BMP-2의 표면 고정화는 불특정 광고를 방지하여 타겟 조사를 허용orption, 성장 인자의 양을 줄임과 동시에, 특히, 표면에서 제어되지 않는 방출을 방해. 세포가 공유 고정 된 rhBMP-2를 제시 표면에 노출되었을 때 BMP-2에 의해 트리거 모두 단기 및 장기의 신호 이벤트는 BMP-2 자극에 대한 세포 반응에 체외 연구를위한이 방법이 적합하다 일어나고있다.

그것은 생물학적으로 불특정 화학적으로 조정할 수있는 시스템을 제공하기 때문에 우리의 설정에서, 금은 부형제로 선정되었습니다. 또한, 자기 조립 단분자막의 응용 프로그램이 많은 이점을 수반한다 : 자신의 기능 최종 그룹이 추가로 수정 될 수있는 반면 SAM에 자발적으로, 금속 및 결함이 거의 형태의 단일 층에 자신의 &quot;머리 그룹&quot;을 통해 흡착. 따라서 그들은 통제 아직 높은 적응력 ?...

Watch this Scientific Journal Video about Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach at JoVE.com

Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

우리는 표면에 BMP-2를 효율적으로 고정화를 달성하기위한 방법을 설명합니다. 우리의 접근은 유리 아민 잔기를 통해 BMP-2 공유 결합을 달성하기 위해 자기 조립 단분자막의 형성에 기초한다. 이 방법은 세포막에서 신호를 연구 할 수있는 유용한 도구입니다.

공유 결합은 자기 조립 단분자막의 접근 방식을 사용하여 표면에 BMP-2의 결합

Bone morphogenetic protein 2 (BMP-2) is a growth factor embedded in the extracellular matrix of bone tissue. BMP-2 acts as trigger of mesenchymal cell ...

covalent-binding-bmp-2-on-surfaces-using-self-assembled-monolayer

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Organic Chemistry

Chemistry

Unit 1

Covalent Bonding and Structure

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13.9K 조회수.  University of Heidelberg.  우리는 표면에 BMP-2를 효율적으로 고정화를 달성하기위한 방법을 설명합니다. 우리의 접근은 유리 아민 잔기를 통해 BMP-2 공유 결합을 달성하기 위해 자기 조립 단분자막의 형성에 기초한다. 이 방법은 세포막에서 신호를 연구 할 수있는 유용한 도구입니다. 

비디오: Covalent Binding of BMP-2 on Surfaces Using a Self-assembled Monolayer Approach

Dependence of Laser-induced Breakdown Spectroscopy Results on Pulse Energies and Timing Parameters Using Soil Simulants

The dependence of some LIBS detection capabilities on lower pulse energies (&lt;100 mJ) and timing parameters were examined using synthetic silicate samples. These samples were used as simulants for soil and contained minor and trace elements commonly found in soil at a wide range of concentrations. For this study, over 100 calibration curves were prepared using different pulse energies and timing parameters; detection limits and sensitivities were determined from the calibration curves. Plasma temperatures were also measured using Boltzmann plots for the various energies and the timing parameters tested. The electron density of the plasma was calculated using the full-width half maximum (FWHM) of the hydrogen line at 656.5 nm over the energies tested. Overall, the results indicate that the use of lower pulse energies and non-gated detection do not seriously compromise the analytical results. These results are very relevant to the design of field- and person-portable LIBS instruments.

LIBS detection capabilities on soil simulants were tested using a range of pulse energies and timing parameters. Calibration curves were used to determine detection limits and sensitivities for different parameters. Generally, the results showed that there was not a significant reduction in detection capabilities using lower pulse energies and non-gated detection.

토양 닮은를 사용하여 펄스 에너지 및 타이밍 매개 변수에 대한 레이저 유도 내역 분광학 결과의 의존

The  dependence of some LIBS detection capabilities on lower pulse energies (&lt;100  mJ) and timing parameters were examined using synthetic silicate ...

연구

화학

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 ...

An Inverse Analysis Approach to the Characterization of Chemical Transport in Paints

The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in understanding contaminant mass transport and the ability to decontaminate materials. If a material is contaminated, over time, the transport of highly toxic chemicals (such as chemical warfare agent species) out of the material can result in vapor exposure or transfer to the skin, which can result in percutaneous exposure to personnel who interact with the material. Due to the high toxicity of chemical warfare agents, the release of trace chemical quantities is of significant concern. Mapping subsurface concentration distribution and transport characteristics of absorbed agents enables exposure hazards to be assessed in untested conditions. Furthermore, these tools can be used to characterize subsurface reaction dynamics to ultimately design improved decontaminants or decontamination procedures. To achieve this goal, an inverse analysis mass transport modeling approach was developed that utilizes time-resolved mass spectroscopy measurements of vapor emission from contaminated paint coatings as the input parameter for calculation of subsurface concentration profiles. Details are provided on sample preparation, including contaminant and material handling, the application of mass spectrometry for the measurement of emitted contaminant vapor, and the implementation of inverse analysis using a physics-based diffusion model to determine transport properties of live chemical warfare agents including distilled mustard (HD) and the nerve agent VX.

In this paper, a procedure for quantifying the mass transport parameters of chemicals in various materials is presented. This process involves employing an inverse-analysis based diffusion model to vapor emission profiles recorded by real-time, mass spectrometry in high vacuum.

페인트의 화학 물질 운송의 특성을 역 분석 접근

The ability to directly characterize chemical transport and interactions that occur within a material (i.e., subsurface dynamics) is a vital component in ...

Dynamic Electrochemical Measurement of Chloride Ions

This protocol describes the dynamic measurement of chloride ions using the transition time of a silver silver chloride (Ag/AgCl) electrode. Silver silver chloride electrode is used extensively for potentiometric measurement of chloride ions concentration in electrolyte. In this measurement, long-term and continuous monitoring is limited due to the inherent drift and the requirement of a stable reference electrode. We utilized the chronopotentiometric approach to minimize drift and avoid the use of a conventional reference electrode. A galvanostatic pulse is applied to an Ag/AgCl electrode which initiates a faradic reaction depleting the Cl&#713; ions near the electrode surface. The transition time, which is the time to completely deplete the ions near the electrode surface, is a function of the ion concentration, given by the Nernst equation. The square root of the transition time is in linear relation to the chloride ion concentration. Drift of the response over two weeks is negligible (59 &#181;M/day) when measuring 1 mM [Cl&#713;]using a current pulse of 10 Am-2. This is a dynamic measurement where the moment of transition time determines the response and thus is independent of the absolute potential. Any metal wire can be used as a pseudo-reference electrode, making this approach feasible for long-term measurement inside concrete structures.

Dynamic measurement of chloride ions is presented. Transition time of an Ag/AgCl electrode, during a chronopotentiometric technique, can give the concentration of chloride ions in electrolyte. This method does not require a stable conventional reference electrode.

This protocol describes the dynamic measurement of chloride ions using the transition time of a silver silver chloride (Ag/AgCl) electrode. Silver silver ...

Facile Preparation of Internally Self-assembled Lipid Particles Stabilized by Carbon Nanotubes

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled (functionalized) CNTs are used as stabilizers to produce Pickering type oil-in-water (O/W) emulsions. Lipids namely, Dimodan U and Phytantriol are used as emulsifiers, which in excess water self-assemble into the bicontinuous cubic Pn3m phase. This highly viscous phase is fragmented into smaller particles using a probe ultrasonicator in presence of conventional surfactant stabilizers or CNTs as done here. Initially, the CNTs (powder form) are dispersed in water followed by further ultrasonication with the molten lipid to form the final emulsion. During this process the CNTs get coated with lipid molecules, which in turn are presumed to surround the lipid droplets to form a particulate emulsion that is stable for months. The average size of CNT-stabilized nanostructured lipid particles is in the submicron range, which compares well with the particles stabilized using conventional surfactants. Small angle X-ray scattering data confirms the retention of the original Pn3m cubic phase in the CNT-stabilized lipid dispersions as compared to the pure lipid phase (bulk state). Blue shift and lowering of the intensities in characteristic G and G&#39; bands of CNTs observed in Raman spectroscopy characterize the interaction between CNT surface and lipid molecules. These results suggest that the interactions between the CNTs and lipids are responsible for their mutual stabilization in aqueous solutions. As the concentrations of CNTs employed for stabilization are very low and lipid molecules are able to functionalize the CNTs, the toxicity of CNTs is expected to be insignificant while their biocompatibility is greatly enhanced. Hence the present approach finds a great potential in various biomedical applications, for instance, for developing hybrid nanocarrier systems for the delivery of multiple functional molecules as in combination therapy or polytherapy.

We report on a smart application of carbon nanotubes for kinetic stabilization of lipid particles that contain self-assembled nanostructures in their cores. The preparation of lipid particles requires rather low concentrations of carbon nanotubes permitting their use in biomedical applications such as drug delivery.

탄소 나노 튜브에 의해 안정화 내부 자기 조립 된 지질 입자의 용이 한 제조

We present a facile method to prepare nanostructured lipid particles stabilized by carbon nanotubes (CNTs). Single-walled (pristine) and multi-walled ...

Highly Stable, Functional Hairy Nanoparticles and Biopolymers from Wood Fibers: Towards Sustainable Nanotechnology

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While metal-based nanoparticles are associated with synthetic and environmental hassles, cellulose introduces a green, sustainable alternative for nanoparticle synthesis. Here, we present the chemical synthesis and separation procedures to produce new classes of hairy nanoparticles (bearing both amorphous and crystalline regions) and biopolymers based on wood fibers. Through periodate oxidation of soft wood pulp, the glucose ring of cellulose is opened at the C2-C3 bond to form 2,3-dialdehyde groups. Further heating of the partially oxidized fibers (e.g., T = 80 &#176;C) results in three products, namely fibrous oxidized cellulose, sterically stabilized nanocrystalline cellulose (SNCC), and dissolved dialdehyde modified cellulose (DAMC), which are well separated by intermittent centrifugation and co-solvent addition. The partially oxidized fibers (without heating) were used as a highly reactive intermediate to react with chlorite for converting almost all aldehyde to carboxyl groups. Co-solvent precipitation and centrifugation resulted in electrosterically stabilized nanocrystalline cellulose (ENCC) and dicarboxylated cellulose (DCC). The aldehyde content of SNCC and consequently surface charge of ENCC (carboxyl content) were precisely controlled by controlling the periodate oxidation reaction time, resulting in highly stable nanoparticles bearing more than 7 mmol functional groups per gram of nanoparticles (e.g., as compared to conventional NCC bearing &#60;&#60; 1 mmol functional group/g). Atomic force microscopy (AFM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) attested to the rod-like morphology. Conductometric titration, Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), dynamic light scattering (DLS), electrokinetic-sonic-amplitude (ESA) and acoustic attenuation spectroscopy shed light on the superior properties of these nanomaterials.

Synthesis schemes to prepare highly stable wood fiber-based hairy nanoparticles and functional cellulose-based biopolymers have been detailed.

매우 안정, 기능 털이 나노 입자와 목재 섬유에서 생체 고분자 : 지속 가능한 나노 기술쪽으로

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While ...

Microfluidic-based Synthesis of Covalent Organic Frameworks (COFs): A Tool for Continuous Production of COF Fibers and Direct Printing on a Surface

Covalent Organic Frameworks (COFs) are a class of porous covalent materials which are frequently synthesized as unprocessable crystalline powders. The first COF was reported in 2005 with much effort centered on the establishment of new synthetic routes for its preparation. To date, most available synthetic methods for COF synthesis are based on bulk mixing under solvothermal conditions. Therefore, there is increasing interest in developing systematic protocols for COF synthesis that provide for fine control over reaction conditions and improve COF processability on surfaces, which is essential for their use in practical applications. Herein, we present a novel microfluidic-based method for COF synthesis where the reaction between two constituent building blocks, 1,3,5-benzenetricarbaldehyde (BTCA) and 1,3,5-tris(4-aminophenyl)benzene (TAPB), takes place under controlled diffusion conditions and at room temperature. Using such an approach yields sponge-like, crystalline fibers of a COF material, hereafter called MF-COF. The mechanical properties of MF-COF and the dynamic nature of the approach allow the continuous production of MF-COF fibers and their direct printing onto surfaces. The general method opens new potential applications requiring advanced printing of 2D or 3D COF structures on flexible or rigid surfaces.

Microfluidic-based Synthesis of Covalent Organic Frameworks COFs: A Tool for Continuous Production of COF Fibers and Direct Printing on a Surface

We present a novel microfluidic-based method for synthesis of covalent organic frameworks (COFs). We demonstrate how this approach can be used to produce continuous COF fibers, and also 2D or 3D COF structures on surfaces.

Microfluidic 기반 CODMs의 합성 : COF 섬유의 지속적인 생산과 표면의 직접 인쇄를위한 도구

Covalent Organic Frameworks (COFs) are a class of porous covalent materials which are frequently synthesized as unprocessable crystalline powders. The ...

Light-driven Molecular Motors on Surfaces for Single Molecular Imaging

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces are demonstrated. This work requires careful design, considerable synthetic effort, and proper analysis. The rotary motion of the molecular motor in solution is shown by 1H NMR and UV-vis absorption spectroscopy techniques. In addition, the method to graft the motor onto an amine-coated quartz is described. This method helps to gain more insight into molecular machines.

The manuscript describes how to synthesize and graft a molecular motor on surfaces for single molecular imaging.

빛 주도 분자 모터 표면에 단일 분자 이미징

The design and synthesis of a synthetic system that aims for the direct visualization of a synthetic rotary motor at the single molecule level on surfaces ...

Ion Mobility-Mass Spectrometry Techniques for Determining the Structure and Mechanisms of Metal Ion Recognition and Redox Activity of Metal Binding Oligopeptides

Electrospray ionization (ESI) can transfer an aqueous-phase peptide or peptide complex to the gas-phase while conserving its mass, overall charge, metal-binding interactions, and conformational shape. Coupling ESI with ion mobility-mass spectrometry (IM-MS) provides an instrumental technique that allows for simultaneous measurement of a peptide&#8217;s mass-to-charge (m/z) and collision cross section (CCS) that relate to its stoichiometry, protonation state, and conformational shape. The overall charge of a peptide complex is controlled by the protonation of 1) the peptide&#8217;s acidic and basic sites and 2) the oxidation state of the metal ion(s). Therefore, the overall charge state of a complex is a function of the pH of the solution that affects the peptides metal ion binding affinity. For ESI-IM-MS analyses, peptide and metal ions solutions are prepared from aqueous-only solutions, with the pH adjusted with dilute aqueous acetic acid or ammonium hydroxide. This allows for pH dependence and metal ion selectivity to be determined for a specific peptide. Furthermore, the m/z and CCS of a peptide complex can be used with B3LYP/LanL2DZ molecular modeling to discern binding sites of the metal ion coordination and tertiary structure of the complex. The results show how ESI-IM-MS can characterize the selective chelating performance of a set of alternative methanobactin peptides and compare them to the copper-binding peptide methanobactin.

Ion mobility-mass spectrometry and molecular modeling techniques can characterize the selective metal chelating performance of designed metal-binding peptides and the copper-binding peptide methanobactin. Developing new classes of metal chelating peptides will help lead to therapeutics for diseases associated with metal ion misbalance.

금속 결합 올리고펩타이드의 금속 이온 인식 및 Redox 활성의 구조 및 메커니즘을 결정하기 위한 이온 이동성 질량 분광법 기술

Electrospray ionization (ESI) can transfer an aqueous-phase peptide or peptide complex to the gas-phase while conserving its mass, overall charge, ...

Synthesis of Information-bearing Peptoids and their Sequence-directed Dynamic Covalent Self-assembly

This protocol presents the use of Lewis acidic multi-role reagents to circumvent kinetic trapping observed during the self-assembly of information-encoded oligomeric strands mediated by paired dynamic covalent interactions in a manner mimicking the thermal cycling commonly employed for the self-assembly of complementary nucleic acid sequences. Primary amine monomers bearing aldehyde and amine pendant moieties are functionalized with orthogonal protecting groups for use as dynamic covalent reactant pairs. Using a modified automated peptide synthesizer, the primary amine monomers are encoded into oligo(peptoid) strands through solid-phase submonomer synthesis. Upon purification by high-performance liquid chromatography (HPLC) and characterization by electrospray ionization mass spectrometry (ESI-MS), sequence-specific oligomers are subjected to high-loading of a Lewis acidic rare-earth metal triflate which both deprotects the aldehyde moieties and affects the reactant pair equilibrium such that strands completely dissociate. Subsequently, a fraction of the Lewis acid is extracted, enabling annealing of complementary sequence-specific strands to form information-encoded molecular ladders characterized by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). The simple procedure outlined in this report circumvents kinetic traps commonly experienced in the field of dynamic covalent assembly and serves as a platform for the future design of robust, complex architectures.

A protocol is presented for the synthesis of information-encoded peptoid oligomers and for the sequence-directed self-assembly of these peptoids into molecular ladders using amines and aldehydes as dynamic covalent reactant pairs and Lewis acidic rare-earth metal triflates as multi-role reagents.

정보 베어링 펩토이드의 합성과 시퀀스 지향 동적 공유자 자체 조립

This protocol presents the use of Lewis acidic multi-role reagents to circumvent kinetic trapping observed during the self-assembly of information-encoded ...