Name: making conjugation-induced fluorescent pegylated virus-like particles by dibromomaleimide-disulfide chemistry
Uploaded: 2018-05-27T14:00:00
Description: Watch this Scientific Journal Video about Making Conjugation-induced Fluorescent PEGylated Virus-like Particles by Dibromomaleimide-disulfide Chemistry at JoVE.com

J.J.G.는 그들의 지원에 대 한 국립 과학 재단 (DMR-1654405) 및 암 예방 연구 연구소의 텍사스 (CPRIT) (RP170752s)을 인정 한다.

1입니다. 준비
<ol><li>Lysogeny 국물 (파운드) 한 천 만들고 접시12부.</li>
	<li>
변환 BL21(DE3) wtQβ 외 투 단백질 시퀀스를 포함 하는 pET28 플라스 미드와.
	<ol>
<li>대장균 BL21(DE3) 유능한 세포를 얼음 목욕에 녹여 Microcentrifuge 튜브에서 셀의 장소 50 μ.</li>
		<li>1 개의 관으로 플라스 미드의 2 μ를 추가 하 고 부드럽게 튜브를 끄 적. 다음 30 분 동안 얼음에 품 어.</li>
		<li>?...

<ol>
	<li>Pokorski, J., Breitenkamp, K., Liepold, L., Qazi, S., Finn, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+Virus-Based+Polymer-Protein+Nanoparticles+by+Atom+Transfer+Radical+Polymerization.">Functional Virus-Based Polymer-Protein Nanoparticles by Atom Transfer Radical Polymerization.</a> J. Am. Chem. Soc. 133 (24), 9242-9245 (2011).</li><li>Capehart, S., Coylet, M., Glasgow, J., Francis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled+Integration+of+Gold+Nanoparticles+and+Organic+Fluorophores+Using+Synthetically+Modified+MS2+Viral+Capsids.">Controlled Integration of Gold Nanoparticles and Organic Fluorophores Using Synthetically Modified MS2 Viral Capsids.</a> J. Am. Chem. Soc. 135 (8), 3011-3016 (2013).</li><li>Li, S., 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=Template-Directed+Synthesis+of+Porous+and+Protective+Core-Shell+Bionanoparticles.">Template-Directed Synthesis of Porous and Protective Core-Shell Bionanoparticles.</a> Angew. Chem. Int. Ed. 55 (36), 10691-10696 (2016).</li><li>Chen, Z., Li, N., Li, S., Dharmarwardana, M., Schlimme, A., Gassensmith, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+Chemistry:+The+Chemical+Functionalization+of+Viral+Architectures+to+Create+New+Technology.">Viral Chemistry: The Chemical Functionalization of Viral Architectures to Create New Technology.</a> WIREs. Nanomed. Nanobiotechnol. 8 (4), 512-534 (2015).</li><li>Chalker, J. M., Bernardes, G. J. L., Lin, Y. A., Davis, B. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+modification+of+proteins+at+cysteine:+opportunities+in+chemistry+and+biology.">Chemical modification of proteins at cysteine: opportunities in chemistry and biology.</a> Chem. - Asian J. 4 (5), 630-640 (2009).</li><li>Le, D. H., Lee, K. L., Shukla, S., Commandeur, U., Steinmetz, N. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potato+Virus+X,+a+Filamentous+Plant+Viral+Nanoparticle+for+Doxorubicin+Delivery+in+Cancer+Therapy.">Potato Virus X, a Filamentous Plant Viral Nanoparticle for Doxorubicin Delivery in Cancer Therapy.</a> Nanoscale. 9 (6), 2348-2357 (2017).</li><li>Chen, L., Wu, Y., Yuan, L., Wang, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virus-templated+FRET+Platform+for+the+Rational+Design+of+Ratiometric+Fluorescent+Nanosensors.">Virus-templated FRET Platform for the Rational Design of Ratiometric Fluorescent Nanosensors.</a> Chem. Comm. 51 (50), 10190-10193 (2015).</li><li>Lee, P., 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=Polymer+Structure+and+Conformation+Alter+the+Antigenicity+of+Virus-like+Particle-Polymer+Conjugates.">Polymer Structure and Conformation Alter the Antigenicity of Virus-like Particle-Polymer Conjugates.</a> J. Am. Chem. Soc. 139 (9), 3312-3315 (2017).</li><li>Zhang, X., 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=Polymer-Protein+Core-Shell+Nanoparticles+for+Enhanced+Antigen+Immunogenicity.">Polymer-Protein Core-Shell Nanoparticles for Enhanced Antigen Immunogenicity.</a> ACS Macro Lett. 6 (4), 442-446 (2017).</li><li>Brown, S. D., Fielder, J. D., Finn, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assembly+of+Hybrid+Bacteriophage+Qbeta+virus-like+particles.">Assembly of Hybrid Bacteriophage Qbeta virus-like particles.</a> Biochemistry. 48 (47), 11155-11157 (2009).</li><li>Chen, Z., 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=Fluorescent+Functionalization+across+Quaternary+Structure+in+a+Virus-+like+Particle.">Fluorescent Functionalization across Quaternary Structure in a Virus- like Particle.</a> Bioconjugate Chem. 28 (9), 2277-2283 (2017).</li><li>. Pouring LB Agar Plates Available from: <a target="_blank" href="https://www.addgene.org/protocols/pouring-lb-agar-plates/">https://www.addgene.org/protocols/pouring-lb-agar-plates/</a> (2016)</li><li>Smith, M., 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=Protein+Modification,+Bioconjugation,+and+Disulfide+Bridging+Using+Bromomaleimides.">Protein Modification, Bioconjugation, and Disulfide Bridging Using Bromomaleimides.</a> J. Am. Chem. Soc. 132 (6), 1960-1965 (2010).</li><li>Castaneda, L., 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=A+Mild+Synthesis+of+N-functionalised+Bromomaleimides,+Thiomaleimides+and+Bromopyridazinediones.">A Mild Synthesis of N-functionalised Bromomaleimides, Thiomaleimides and Bromopyridazinediones.</a> Tetrahedron Lett. 54 (27), 3493-3495 (2013).</li><li>Fiedler, J., 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=Engineered+Mutations+Change+the+Structure+and+Stability+of+a+Virus-+Like+Particle.">Engineered Mutations Change the Structure and Stability of a Virus- Like Particle.</a> Biomacromolecules. 13 (8), 2339-2348 (2012).</li><li>Manzenrieder, F., Luxenhofer, R., Retzlaff, M., Jordan, R., Finn, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stabilization+of+Virus-like+Particles+with+Poly(2-oxazoline)s.">Stabilization of Virus-like Particles with Poly(2-oxazoline)s.</a> Angew. Chem. Int. Ed. 50 (11), 2601-2605 (2011).</li><li>Chen, Z., Li, N., Chen, L., Lee, J., Gassensmith, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual+Functionalized+Bacteriophage+Q&#946;+as+a+Photocaged+Drug+Carrier.">Dual Functionalized Bacteriophage Q&#946; as a Photocaged Drug Carrier.</a> Small. 12 (33), 4563-4571 (2016).</li></ol>

작은 단백질 정화에 비해, 살 균 소 Qβ 정화는 독특한 단계 자당 기온 변화도 원심 분리입니다. 클로 프롬/n-butanol 추출 단계 후 Qβ 5-40% 자당 기온 변화도 사용 하 여 더 순화 이다. 원심 분리, 중 입자는 그들의 크기에 따라 구분 됩니다. 더 큰 입자 작은 입자는 낮은 밀도 영역에 체류 하는 동안 더 높은 밀도 영역에 여행. Qβ는 그라데이션의 더 낮은 세 번째에 여행 하 고 더 작은 단백질 불순물은 원...

바이러스 성 capsids 나노 크기를 사용 하 여 생물 의학 연구1,2,3응용 프로그램의 범위를 확대 하는 것을 목표로 하는 흥미 진 진한 필드로 떠오르고 있다. Recombinantly 표현된 바이러스 같은 입자 (VLPs)는 구조적으로 바이러스에서 파생 된 하지만 그들은 그들에 게 비 전염 성 배치할 나노 입자를 만드는 원래 바이러스 유전 물질 부족 합니다. 표면 기능 프로그램 유전자는 고 각 capsid 앞뒤 사람에 게 동일 하 게 표현 된다, 위치 및 원자 정밀도로 아미노산의 반응 측면 체인의 수를 알아야 가능 하다. 대부분의 경우에서 외부와 실내 표면 bioconjugation 반응-는 biomolecule와 합성 사이 공유 결합을 형성 하는 반응을 통해 functionalized feasibly 수 있습니다 용 매 노출된 아미노산 잔류물의 많은 종류를가지고 분자4,5.
Bioconjugation 반응 생체의 비교적 간단한 패션에서 더 다양 한 기능을가지고 도움이 됩니다. 분자의, 치료 약6, 형광 태그7 등 폴리머8,9 미리 합성 하 고 특징 VLPs의 표면에 연결 하기 전에 수 있습니다. 생물 의학에서 특히 일반적인 VLP 바이오 소재 연구, recombinantly 표현으로 28 nm icosahedral 바이러스 성 capsid10살 균 소 Qβ에 따라 VLP 되었습니다. 비록 우리가 최근에 Haddleton 베이커 반응을 통해 Qβ의 숨 구멍 줄 감소 disulfides dibromomaleimide 화합물의 성공적인 활용11 전달 Qβ에 가장 일반적인 반응 사이트 넓은 여백에 의해 lysines 있습니다. 반응 좋은 진행 항복 하 고, 동등 하 게 중요 한 것은, 입자의 열 안정성을 잃고 없이. 동시에이 반응을 활용 유도 형광, 셀으로이 입자의 통풍 관을 추적 하는 데 사용할 수 있습니다 생성 합니다. 이 작품에서는, 우리 Haddleton 베이커 반응, 밝은 노란색 fluorophore를 통해 Qβ의 표면에 폴 리 에틸렌 글리콜 (PEG)의 활용을 보여 줍니다. 이러한 입자 세포에 의해 그들은 촬영으로 추적할 수 있습니다. 본 프로토콜 연구원 원리 용 노출된 disulfides 포함 된 많은 다른 VLPs 중 하나에 적용 되는 새로운 형광 PEGylated 배치할 나노 입자를 기반으로 Qβ, 생성 도움이 됩니다.

<table border="0" cellpadding="0" cellspacing="0" width="573">
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			<td style="width:119px;">S271-10</td>
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			<td height="42" style="height:42px;width:216px;">Potassium Chloride</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">BP366-500</td>
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			<td style="width:119px;">P288-500</td>
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			<td height="42" style="height:42px;width:216px;">Isopropyl &#946;-D-1-thiogalactopyranoside (IPTG)</td>
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			<td height="42" style="height:42px;width:216px;">1-Butanol</td>
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			<td height="84" style="height:84px;width:216px;">SW 28 Ti Rotor, Swinging Bucket, Aluminum</td>
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			<td height="42" style="height:42px;width:216px;">Type 70 Ti Rotor, Fixed Angle, Titanium, 8 x 39 mL,</td>
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			<td style="width:119px;">337922</td>
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			<td height="42" style="height:42px;width:216px;">Coomassie (Bradford) Protein Assay</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">PI23200</td>
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			<td height="84" style="height:84px;width:216px;">TRIS Hydrochloride</td>
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			<td style="width:119px;">T60050-1000.0</td>
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			<td height="42" style="height:42px;width:216px;">Tris(2-carboxyethyl)phosphine hydrochloride</td>
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			<td style="width:119px;">AC424375000</td>
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			<td style="width:119px;">L3771-100G</td>
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			<td height="42" style="height:42px;width:216px;">Ammonium Persulfate</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">BP179-100</td>
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			<td height="42" style="height:42px;width:216px;">FV3000 confocal laser scanning microscope</td>
			<td style="width:71px;">Olympus&#160;</td>
			<td>FV3000&#160;</td>
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			<td height="42" style="height:42px;width:216px;">Labnet Revolver Adjustable Rotator&#160;</td>
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			<td>1190P25&#160;</td>
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			<td style="width:119px;">010-1459</td>
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			<td style="width:119px;">3141-0250</td>
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			<td style="width:71px;">Thermo Scientific</td>
			<td style="width:119px;">3117-0380</td>
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			<td height="42" style="height:42px;width:216px;">2 L Narrow Mouth Erlenmeyer Flasks with Heavy Duty Rim</td>
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			<td style="width:119px;">4980-2L</td>
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making conjugation-induced fluorescent pegylated virus-like particles by dibromomaleimide-disulfide chemistry

바이러스 같은 입자 (VLPs) 생물 의학에 있는 최근 증가 및 재료 연구 생 합성, 개별 크기, 유전자 프로그래밍 및 biodegradability의 그들의 용이 함에 표시 될 수 있습니다. 그들은 그들의 표면에 합성 ligands를 추가 하기 위한 bioconjugation 반응에 매우 의무가 있어, 이러한 수성 태어난된 capsids에 bioconjugation 방법론에서 범위 상대적으로 제한 됩니다. 촉진 하기 위해 기능성 생체 재료 연구의 방향, 비 전통적인 bioconjugation 반응은 고려 되어야 한다. 이 프로토콜에서 설명 하는 반응 dibromomaleimides를 사용 하 여 살 균 소 Qβ를 기반으로 한 VLP의 노출된 이황화 결합 용 매에서 새로운 기능을 소개. 또한, 최종 제품은 형광, 추적이 가능한 생체 외에서 프로브를 상업적으로 사용할 수 있는 필터 세트를 사용 하 여 생성의 이점이 있다 이다.

Dibromomaleimide 파생 상품 dibromomaleimide 무수 물 및 1 차 아민15사이 응축 반응을 통해 합성 될 수 있다. 또는 N-methoxycarbonyl 활성화 3, 4-dibromomaleimide를 사용 하 여 가벼운 합성 방법16 methoxypolyethylene 글리콜 (PEG) 수익률 DB-페그 (그림 1)에 반응 하 여 여기 악용 했다. NMR 화합물 구조 (그림 2)를 식별 하?...

Watch this Scientific Journal Video about Making Conjugation-induced Fluorescent PEGylated Virus-like Particles by Dibromomaleimide-disulfide Chemistry at JoVE.com

Making Conjugation-induced Fluorescent PEGylated Virus-like Particles by Dibromomaleimide-disulfide Chemistry

Dibromomaleimide-아 황산 화학에 의해 형광 PEGylated 바이러스 같은 입자 활용 유도 만들기

여기, 선물이 붙일 functionalize dibromomaleimide와 Qβ VLP에 disulfides에 절차. 우리는 Qβ 식 및 정화, dibromomaleimide 기능성된 분자의 합성 및 dibromomaleimide와 Qβ 사이 활용 반응 설명합니다. 결과 노란색 형광 활용된 입자 셀 안에 형광 프로브로 사용할 수 있습니다.

The recent rise in virus-like particles (VLPs) in biomedical and materials research can be attributed to their ease of biosynthesis, discrete size, ...

Research

JoVE Journal

Bioengineering

A Microfluidic-based Hydrodynamic Trap for Single Particles

단일 입자 Microfluidic 기반 유체 트랩

The ability to confine and manipulate single particles in free solution is a key enabling technology for fundamental and applied science.  Methods for ...

연구

생체공학

Thermodynamics of Membrane Protein Folding Measured by Fluorescence Spectroscopy

막 단백질 폴딩의 열역학은 형광 분광학에 의해 측정

Membrane protein folding is an emerging topic with both fundamental and health-related significance.  The abundance of membrane proteins in cells ...

Imaging of Biological Tissues by Desorption Electrospray Ionization Mass Spectrometry

탈착 전기 분무 이온화 질량 분석에 의한 생물 조직의 이미지

Mass spectrometry imaging (MSI) provides untargeted molecular information with the highest specificity and spatial resolution for investigating biological ...

Intra-lymph Node Injection of Biodegradable Polymer Particles

생분해성 폴리머 입자의 림프절 내 주입

Generation of adaptive immune response relies on efficient drainage or trafficking of antigen to lymph nodes for processing and presentation of these ...

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

마이크로 RNA 정량는 DNA 결합 염료 화학, 물방울 디지털 PCR을 사용하여 순환

Circulating (of cell-free) microRNAs (miRNAs) are released from cells into the blood stream. The amount of specific microRNAs in the circulation has been ...

Determination of Zeta Potential via Nanoparticle Translocation Velocities through a Tunable Nanopore: Using DNA-modified Particles as an Example

조정 나노 기공을 통해 나노 입자 전좌 속도를 통해 제타 전위의 측정 : 예를 들어 DNA 변성 입자를 사용하여

Nanopore technologies, known collectively as Resistive Pulse Sensors (RPS), are being used to detect, quantify and characterize proteins, molecules and ...

Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles

입자의 공간 추적을위한 멀티 플렉스 전자 감지와 미세 유체 플랫폼

Microfluidic processing of biological samples typically involves differential manipulations of suspended particles under various force fields in order to ...

Trapping of Micro Particles in Nanoplasmonic Optical Lattice

Nanoplasmonic 광학 격자에 마이크로 입자의 트래핑

The plasmonic optical tweezer has been developed to overcome the diffraction limits of the conventional far field optical tweezer. Plasmonic optical ...

Site-Directed Immobilization of Bone Morphogenetic Protein 2 to Solid Surfaces by Click Chemistry

뼈 Morphogenetic 단백질의 2 고체 표면에 화학 클릭 하 여 사이트 감독 동원 정지

Different therapeutic strategies for the treatment of non-healing long bone defects have been intensively investigated. Currently used treatments present ...

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence