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专注多肽 服务科研
"Peptide H-AHIVMVDAYKPTK-OH is a Research Peptide with significant interest within the field academic and medical research. Recent citations using H-AHIVMVDAYKPTK-OH include the following: Engineering tunable dual functional protein cage nanoparticles using bacterial superglue Y Bae, GJ Kim, H Kim , SG Park, HS Jung - , 2018 - ACS Publicationshttps://pubs.acs.org/doi/abs/10.1021/acs.biomac.8b00457 Development of Functional Protein-based Target-specific Labeling Nanoplatforms for Biological Applications Y Bae - 2021 - scholarworks.unist.ac.krhttps://scholarworks.unist.ac.kr/bitstream/201301/50394/1/Development%20of%20Functional%20Protein-based%20Target-specific%20Labeling%20Nanoplatforms%20for%20Biological%20Applications.pdf Virus-like particle display of HER2 induces potent anti-cancer responses VG Ianzano, TG Theander , MA Nielsen , A Salanti - academia.eduhttps://www.academia.edu/download/81646784/2162402X.2017.pdf Spying on neuronal membrane potential with genetically targetable voltage indicators V Grenier , BR Daws, P Liu - Journal of the American , 2019 - ACS Publicationshttps://pubs.acs.org/doi/abs/10.1021/jacs.8b11997 Improving the malaria transmission-blocking activity of a Plasmodium falciparum 48/45 based vaccine antigen by SpyTag/SpyCatcher mediated virus-like display SK Singh, S Thrane, CM Janitzek, MA Nielsen - Vaccine, 2017 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0264410X17306904 Pfs230 and Pfs48/45 Fusion Proteins Elicit Strong Transmission-Blocking Antibody Responses Against Plasmodium falciparum SK Singh, S Thrane , BK Chourasia , K Teelen - Frontiers in , 2019 - frontiersin.orghttps://www.frontiersin.org/articles/10.3389/fimmu.2019.01256/full Enzymatic bioconjugation of nanoparticles: developing specificity and control SA Walper , KB Turner, IL Medintz - Current Opinion in Biotechnology, 2015 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S095816691500066X Bacterial superglue enables easy development of efficient virus-like particle based vaccines S Thrane, CM Janitzek, S Matondo, M Resende - Journal of , 2016 - Springerhttps://link.springer.com/article/10.1186/s12951-016-0181-1 Ambient temperature stable, scalable COVID-19 polymer particle vaccines induce protective immunity S Chen , B Evert, A Adeniyi - Advanced , 2022 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1002/adhm.202102089 Site-Specific Fluorescent Labeling of Antibodies and Diabodies Using SpyTag/SpyCatcher System for In Vivo Optical Imaging MK Alam , A El-Sayed , K Barreto , W Bernhard - Molecular imaging and , 2019 - Springerhttps://link.springer.com/article/10.1007/s11307-018-1222-y Engineered hepatitis B core virus-like particle carrier for precise and personalized Alzheimers disease vaccine preparation via fixed-point coupling M Ji, X Xie, D Liu, S Lu, L Zhang , Y Huang, R Liu - Applied Materials Today, 2020 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S2352940720300226 Spontaneous and site-specific immobilization of PNGase F via spy chemistry L Zhang, W Wang, Y Yang, X Liu, W Zhu, L Pi, X Liu - RSC , 2023 - pubs.rsc.orghttps://pubs.rsc.org/en/content/articlehtml/2023/ra/d3ra04591a Thermostable T Cell Multiepitope Nanoparticle Antigens Inducing Potent Immune Responses against the Swine Fever Virus L Sun, J Niu, J Zhang, Y Peng, X Feng - ACS Infectious , 2023 - ACS Publicationshttps://pubs.acs.org/doi/abs/10.1021/acsinfecdis.3c00506 Structural analysis and optimization of the covalent association between SpyCatcher and a peptide Tag L Li , JO Fierer, TA Rapoport, M Howarth - Journal of molecular biology, 2014 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0022283613006669 Non-neutralizing epitopes shade neutralizing epitopes against omicron in a multiple epitope-based vaccine HR Gong, Y Hu , X Li, T Yau , BZ Zhang - ACS Infectious , 2022 - ACS Publicationshttps://pubs.acs.org/doi/abs/10.1021/acsinfecdis.2c00488 Creation of a formate: malate oxidoreductase by fusion of dehydrogenase enzymes with PEGylated cofactor swing arms HF Ozbakir , KE Garcia, S Banta - Protein Engineering, Design , 2018 - academic.oup.comhttps://academic.oup.com/peds/article-abstract/31/4/103/4962187 Tumor targetable high-field MRI T1 contrast conjugates with Gd (III)-chelate complexes and protein cage nanoparticles H Kim, S Jin, H Choi, MS Kang, Y Bae, HJ Cho , S Kang - abstracts.biomaterials.orghttp://abstracts.biomaterials.org/data/papers/2019/abstracts/989.pdf Target-switchable Gd (III)-DOTA/protein cage nanoparticle conjugates with multiple targeting affibody molecules as target selective T1 contrast agents for high-field H Kim , S Jin, H Choi, MS Kang, SG Park, H Jun - Journal of Controlled , 2021 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0168365921002534 DNA-Encoded Multivalent Display of Chemically Modified Protein Tetramers on Phage: Synthesis and in Vivo Applications GM Lima , A Atrazhev, S Sarkar , M Sojitra - ACS Chemical , 2021 - ACS Publicationshttps://pubs.acs.org/doi/abs/10.1021/acschembio.1c00835 Resurfaced ZIKV EDIII nanoparticle immunogens elicit neutralizing and protective responses in vivo GI Georgiev, RJ Malonis , AS Wirchnianski - Cell chemical , 2022 - cell.comhttps://www.cell.com/cell-chemical-biology/pdf/S2451-9456(22)00085-X.pdf A nanoparticle vaccine displaying the ookinete PSOP25 antigen elicits transmission-blocking antibody response against Plasmodium berghei G Yao, H Min , X Yu, F Liu , L Cui , Y Cao - Parasites & Vectors, 2023 - Springerhttps://link.springer.com/article/10.1186/s13071-023-06020-8 Methods and applications D Min, MA Arbing , RE Jefferson, JU Bowie - researchgate.nethttps://www.researchgate.net/profile/Mark-Arbing/publication/303510516_A_simple_DNA_handle_attachment_method_for_single_molecule_mechanical_manipulation_experiments/links/6356a05c6e0d367d91c0a37d/A-simple-DNA-handle-attachment-method-for-single-molecule-mechanical-manipulation-experiments.pdf The immunogenicity of capsid-like particle vaccines in combination with different adjuvants using different routes of administration CM Janitzek, PHR Carlsen, S Thrane , VM Khanna - Vaccines, 2021 - mdpi.comhttps://www.mdpi.com/2076-393X/9/2/131 Spying on Neuronal B Daws, P Liu, E Miller - 2018 - scholar.archive.orghttps://scholar.archive.org/work/fzg3y6mhgnhybeigfokn6727gi/access/wayback/https://s3-eu-west-1.amazonaws.com/itempdf74155353254prod/7313657/Spying_on_Neuronal_Membrane_Potential_with_Genetically_Targetable_Voltage_Indicators_v1.pdf Fabrication of modularly functionalizable microcapsules using protein-based technologies AC Schloss, W Liu , DM Williams - ACS biomaterials , 2016 - ACS Publicationshttps://pubs.acs.org/doi/abs/10.1021/acsbiomaterials.6b00447 Molecular Tags for Proteins and Their Biological Applications A Basak , S Basak - Current Proteomics, 2018 - ingentaconnect.comhttps://www.ingentaconnect.com/content/ben/cp/2018/00000015/00000005/art00010"
专肽生物可以提供常见抗体制备、免疫学研究等生物科研需求的各类标签多肽(Tag peptide)以及蛋白底物产品,可用荧光标记(FITC、5-Fam)等。除常见抗体中抗原决定部位的标签多肽(tag: His | c-Myc | HA | FLAG | V5 | X-press | VSV tag | T7 tag | Snoop tag | E-tag | Sof tag 1 | Spy tag |等等),我们还提供各类定制标签肽的合成,并可提供荧光标记(FITC、5-Fam)等。同时,免费提供的MS和HPLC分析报告。
多肽H2N-Ala-His-Ile-Val-Met-Val-Asp-Ala-Tyr-Lys-Pro-Thr-Lys-COOH的合成步骤:
1、合成CTC树脂:称取2.9g CTC Resin(如初始取代度约为0.4mmol/g)和1.39mmol Fmoc-Lys(Boc)-OH于反应器中,加入适量DCM溶解氨基酸(需要注意,此时CTC树脂体积会增大好几倍,避免DCM溶液过少),再加入3.48mmol DIPEA(Mw:129.1,d:0.740g/ml),反应2-3小时后,可不抽滤溶液,直接加入1ml的HPLC级甲醇,封端半小时。依次用DMF洗涤2次,甲醇洗涤1次,DCM洗涤一次,甲醇洗涤一次,DCM洗涤一次,DMF洗涤2次(这里使用甲醇和DCM交替洗涤,是为了更好地去除其他溶质,有利于后续反应)。得到 Fmoc-Lys(Boc)-CTC Resin。结构图如下:
2、脱Fmoc:加3倍树脂体积的20%Pip/DMF溶液,鼓氮气30分钟,然后2倍树脂体积的DMF 洗涤5次。得到 H2N-Lys(Boc)-CTC Resin 。(此步骤脱除Fmoc基团,茚三酮检测为蓝色,Pip为哌啶)。结构图如下:
3、缩合:取3.48mmol Fmoc-Thr(tBu)-OH 氨基酸,加入到上述树脂里,加适当DMF溶解氨基酸,再依次加入6.96mmol DIPEA,3.31mmol HBTU。反应30分钟后,取小样洗涤,茚三酮检测为无色。用2倍树脂体积的DMF 洗涤3次树脂。(洗涤树脂,去掉残留溶剂,为下一步反应做准备)。得到Fmoc-Thr(tBu)-Lys(Boc)-CTC Resin。氨基酸:DIPEA:HBTU:树脂=3:6:2.85:1(摩尔比)。结构图如下:
4、依次循环步骤二、步骤三,依次得到
H2N-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Ile-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-Ile-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-His(Trt)-Ile-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
H2N-His(Trt)-Ile-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
Fmoc-Ala-His(Trt)-Ile-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin
以上中间结构,均可在专肽生物多肽计算器-多肽结构计算器中,一键画出。
最后再经过步骤二得到 H2N-Ala-His(Trt)-Ile-Val-Met-Val-Asp(OtBu)-Ala-Tyr(tBu)-Lys(Boc)-Pro-Thr(tBu)-Lys(Boc)-CTC Resin,结构如下:
5、切割:6倍树脂体积的切割液(或每1g树脂加8ml左右的切割液),摇床摇晃 2小时,过滤掉树脂,用冰无水乙醚沉淀滤液,并用冰无水乙醚洗涤沉淀物3次,最后将沉淀物放真空干燥釜中,常温干燥24小试,得到粗品H2N-Ala-His-Ile-Val-Met-Val-Asp-Ala-Tyr-Lys-Pro-Thr-Lys-COOH。结构图见产品结构图。
切割液选择:1)TFA:H2O=95%:5%
2)TFA:H2O:TIS=95%:2.5%:2.5%
3)三氟乙酸:茴香硫醚:1,2-乙二硫醇:苯酚:水=87.5%:5%:2.5%:2.5%:2.5%
(前两种适合没有容易氧化的氨基酸,例如Trp、Cys、Met。第三种适合几乎所有的序列。)
6、纯化冻干:使用液相色谱纯化,收集目标峰液体,进行冻干,获得蓬松的粉末状固体多肽。不过这时要取小样复测下纯度 是否目标纯度。
7、最后总结:
杭州专肽生物技术有限公司(ALLPEPTIDE https://www.allpeptide.com)主营定制多肽合成业务,提供各类长肽,短肽,环肽,提供各类修饰肽,如:荧光标记修饰(CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等),功能基团修饰肽(叠氮、炔基、DBCO、DOTA、NOTA等),同位素标记肽(N15、C13),订书肽(Stapled Peptide),脂肪酸修饰肽(Pal、Myr、Ste),磷酸化修饰肽(P-Ser、P-Thr、P-Tyr),环肽(酰胺键环肽、一对或者多对二硫键环),生物素标记肽,PEG修饰肽,甲基化修饰肽等。
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