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OVA Peptide(257-264) 是 OVA 的 I 类 (Kb)-限制性肽表位，OVA 是由 I 类 MHC 分子 H-2Kb 呈递的来自卵清蛋白的八聚体肽。
OVA Peptide(257-264) is a class I (Kb)-restricted peptide epitope of OVA, an octameric peptide from ovalbumin presented by the class I MHC molecule, H-2Kb.

OVA肽（257-264）TFA是OVA的I类（Kb）限制性肽表位，OVA是由I类MHC分子H-2Kb呈递的来自卵清蛋白的八聚体肽。OVA肽（257-264）表位对TAP转运蛋白具有差异依赖性，这取决于OVA表位的蛋白质环境：包含在MBPCrl OVA或Crl OVA细菌融合蛋白中的OVA肽，而全长卵清蛋白中所含的OVA肽（257-264）在很大程度上依赖于TAP转运蛋白，无论重组OVA是在细菌中表达还是天然蛋白与聚苯乙烯珠偶联。

OVA Peptide (257-264) TFA is a class I (Kb)-restricted peptide epitope of OVA, an octameric peptide from ovalbumin presented by the class I MHC molecule, H-2Kb. OVA Peptide (257-264) epitope is presented with a differential dependence on the TAP transporter depending on the protein context of the OVA epitope: OVA Peptide (257-264) contained within the MBPCrl-OVA or Crl-OVA bacterial fusion proteins is presented with little dependence on the TAP transporter, while OVA Peptide (257-264) contained within full-length ovalbumin is largely dependent on the TAP transporter, regardless of whether recombinant OVA is expressed in bacteria or the native protein is coupled to polystyrene beads.

Ovalbumin (257-264) is an acetate salt of a fragment of the protein ovalbumin.

SIINFEKL sequence.OVA Peptide (257-264) is a fragment of the OVA protein that stimulates an antibody response. It has been shown to activate various types of immune cells, such as T-cells, B-cells and macrophages.

Acetate salt. OVA 257-264 (H-2Kb) is an epitope of ovalbumin.

Ovalbumin (OVA) is the primary protein in egg-white, and is involved in initiating food allergies and asthma. It is a highly immunogenic protein and can be used for peptide conjugation in the development of antibodies.OVA (257-264) is a class I (Kb)-restricted peptide epitope of OVA. The ovalbumin fragment is presented by the class I MHC molecule, H-2Kb. Ovalbumin (257-264) chicken has been used to trigger T cell activation in immunology studies.\nSold as the TFA salt

Peptide H-SIINFEKL^-OH is a Research Peptide with significant interest within the field academic and medical research. Recent citations using H-SIINFEKL^-OH include the following: Endogenous viral antigen processing generates peptide-specific MHC class I cell-surface clusters X Lu, JS Gibbs, HD Hickman , A David - Proceedings of the , 2012 - National Acad Scienceshttps://www.pnas.org/doi/abs/10.1073/pnas.1208696109 Dendritic cell-mediated cross presentation of tumor-derived peptides is biased against plasma membrane proteins TB Fessenden , LE Stopfer , F Chatterjee - for ImmunoTherapy of , 2022 - ncbi.nlm.nih.govhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9277371/ Mouse urinary peptides provide a molecular basis for genotype discrimination by nasal sensory neurons T Sturm, T Leinders-Zufall , B MaÃ\x84Â\x8dek , M Walzer - Nature , 2013 - nature.comhttps://www.nature.com/articles/ncomms2610 Short peptide sequences containing MHC class I and/or class II epitopes linked to nano-beads induce strong immunity and inhibition of growth of antigen-specific T Fifis, P Mottram, V Bogdanoska, J Hanley - Vaccine, 2004 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0264410X0400427X of peptide-specific CTL activity and inhibition of tumor growth following immunization with nanoparticles coated with tumor peptide-MHC-I complexes SH Kim, HE Park, SU Jeong, JH Moon, YR Lee - Immune , 2021 - ncbi.nlm.nih.govhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8733191/ A modular antigen presenting peptide/oligonucleotide nanostructure platform for inducing potent immune response S Tohumeken , N Gunduz , MB Demircan - Advanced , 2017 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1002/adbi.201700015 Cutting edge: N-hydroxy peptides: a new class of TCR antagonists S Hin, C Zabel, A Bianco , G Jung - The Journal of , 1999 - journals.aai.orghttps://journals.aai.org/jimmunol/article/163/5/2363/32894 Processing of exogenous antigens for presentation by class I MHC molecules involves post-Golgi peptide exchange influenced by peptide-MHC complex stability and PJ Chefalo, CV Harding - The Journal of Immunology, 2001 - journals.aai.orghttps://journals.aai.org/jimmunol/article/167/3/1274/35053 26S proteasomes and immunoproteasomes produce mainly N-extended versions of an antigenic peptide P Cascio, C Hilton , AF Kisselev, KL Rock - The EMBO , 2001 - embopress.orghttps://www.embopress.org/doi/full/10.1093/emboj/20.10.2357 Exact prediction of a natural T cell epitope O Rötzschke , K Falk, S Stevanovic - European journal of , 1991 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1002/eji.1830211136 Immune evasion proteins of murine cytomegalovirus preferentially affect cell surface display of recently generated peptide presentation complexes NAW Lemmermann , K Gergely, V BoÃ\x8cË\x86hm - Journal of , 2010 - Am Soc Microbiolhttps://journals.asm.org/doi/abs/10.1128/jvi.02087-09 Ovalbumin epitope SIINFEKL self-assembles into a supramolecular hydrogel M Kamalov , H Kahlig, C Rentenberger, ARM Mullner - Scientific Reports, 2019 - nature.comhttps://www.nature.com/articles/s41598-019-39148-8 Tapasin enhances MHC class I peptide presentation according to peptide half-life M Howarth , A Williams, AB Tolstrup - Proceedings of the , 2004 - National Acad Scienceshttps://www.pnas.org/doi/abs/10.1073/pnas.0306294101 Proteasomes and antigen presentation: evidence that a KEKE motif does not promote presentation of the class I epitope SIINFEKL M Gonciarz-Swiatek, M Rechsteiner - Molecular immunology, 2006 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0161589005004104 Isolation of processed, H-2Kb -binding ovalbumin-derived peptides associated with the stress proteins HSP70 and GP96 M Breloer , T Marti, B Fleischer - European journal of , 1998 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1002/(SICI)1521-4141(199803)28:03%3C1016::AID-IMMU1016%3E3.0.CO;2-G Sortase-mediated modification of alphaDEC205 affords optimization of antigen presentation and immunization against a set of viral epitopes LK Swee, CP Guimaraes, S Sehrawat - Proceedings of the , 2013 - National Acad Scienceshttps://www.pnas.org/doi/abs/10.1073/pnas.1214994110 Investigation of antigen delivery route in vivo and immune-boosting effects mediated by pH-sensitive liposomes encapsulated with Kb-restricted CTL epitope KY Lee, E Chun , BL Seong - Biochemical and biophysical research , 2002 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0006291X02967112 Both human and mouse cells expressing H-2Kb and ovalbumin process the same peptide, SIINFEKL K Falk, O Rötzschke , S Faath, S Goth, I Graef - Cellular , 1993 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0008874983712128 MHC/peptide binding studies indicate hierarchy of anchor residues K Deres, W Beck, S Faath, G Jung, HG Rammensee - Cellular immunology, 1993 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0008874983712281 Interferon-γ can stimulate post-proteasomal trimming of the N terminus of an antigenic peptide by inducing leucine aminopeptidase J Beninga, KL Rock, AL Goldberg - Journal of Biological Chemistry, 1998 - ASBMBhttps://www.jbc.org/article/S0021-9258(18)80196-X/abstract A 16-mer peptide (RQIKIWFQNRRMKWKK) from antennapedia preferentially targets the Class I pathway GA Pietersz, W Li, V Apostolopoulos - Vaccine, 2001 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0264410X0000373X Detection of Antigen Presentation by Murine Bone Marrow-Derived Dendritic Cells After Treatment with Nanoparticles E Hong, MA Dobrovolskaia - of Nanoparticles Intended for Drug Delivery, 2024 - Springerhttps://link.springer.com/protocol/10.1007/978-1-0716-3786-9_17 Penetratin tandemly linked to a CTL peptide induces anti-tumour T-cell responses via a cross-presentation pathway DS Pouniotis, V Apostolopoulos , GA Pietersz - Immunology, 2006 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1111/j.1365-2567.2005.02304.x Comparison of protein and peptide targeting for the development of a CD169-based vaccination strategy against melanoma D Van Dinther , H Veninga, M Revet - Frontiers in , 2018 - frontiersin.orghttps://www.frontiersin.org/articles/10.3389/fimmu.2018.01997/full Generation of both MHC class I-and class II-restricted antigenic peptides from exogenously added ovalbumin in murine phagosomes D Muno, E Kominami, T Mizuochi - FEBS letters, 2000 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0014579300018494 A SIINFEKL-based system to measure MHC class I antigen presentation efficiency and kinetics D Dersh , JW Yewdell , J Wei - Antigen Processing: Methods and Protocols, 2019 - Springerhttps://link.springer.com/protocol/10.1007/978-1-4939-9450-2_9 Defective ribosomal products are the major source of antigenic peptides endogenously generated from influenza A virus neuraminidase BP Dolan, L Li, K Takeda, JR Bennink - The journal of , 2010 - journals.aai.orghttps://journals.aai.org/jimmunol/article/184/3/1419/82278 Sulfated Lactosyl Archaeol Archaeosomes Synergize with Poly (I: C) to Enhance the Immunogenicity and Efficacy of a Synthetic Long Peptide-Based Vaccine in B Akache, G Agbayani, FC Stark , Y Jia, R Dudani - 2021 - academia.eduhttps://www.academia.edu/download/79316597/pdf.pdf T cell determinants incorporating beta-amino acid residues are protease resistant and remain immunogenic in vivo AI Webb , MA Dunstone , NA Williamson - The Journal of , 2005 - journals.aai.orghttps://journals.aai.org/jimmunol/article/175/6/3810/73297 Functional screening of a retroviral peptide library for MHC class I presentation AB Tolstrup, M Duch, I Dalum, FS Pedersen - Gene, 2001 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0378111900005722 Peptide-loaded dendritic cells prime and activate MHC-class I-restricted T cells more efficiently than protein-loaded cross-presenting DC acaâ\x80\x93 Met, S Buus , MH Claesson - Cellular immunology, 2003 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S000887490300128X for a novel luminal endoplasmic reticulum aminopeptidase in final trimming of 26 S proteasome-generated major histocompatability complex class I antigenic peptides A Komlosh, F Momburg , T Weinschenk - Journal of Biological , 2001 - ASBMBhttps://www.jbc.org/article/S0021-9258(20)89708-7/abstract pDNA-lipoplexes engrafted with flagellin-related peptide induce potent immunity and anti-tumour effects A Faham, T Herringson, C Parish, A Suhrbier - Vaccine, 2011 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0264410X11010772 Two distinct proteolytic processes in the generation of a major histocompatibility complex class I-presented peptide A Craiu, T Akopian, A Goldberg - Proceedings of the , 1997 - National Acad Scienceshttps://www.pnas.org/doi/abs/10.1073/pnas.94.20.10850 Analysis of the proteolytic steps involved in MHC class I presentation of the ovalbumin epitope SIINFEKL A Craiu - 1997 - search.proquest.comhttps://search.proquest.com/openview/dc58025ab6af04f400622af0d7a743d2/1?pq-origsite=gscholar&cbl=18750&diss=y N-hydroxy-amide analogues of MHC-class I peptide ligands with nanomolar binding affinities A Bianco , C Zabel, P Walden - of the European Peptide , 1998 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1002/(SICI)1099-1387(199812)4:8%3C471::AID-PSC166%3E3.0.CO;2-8 New synthetic non-peptide ligands for classical major histocompatibility complex class I molecules A Bianco , C Brock, C Zabel, T Walk, P Walden - Journal of Biological , 1998 - ASBMBhttps://www.jbc.org/article/S0021-9258(19)59453-4/abstract

多肽H2N-Ser-Ile-Ile-Asn-Phe-Glu-Lys-Leu-COOH的合成步骤：

1、合成CTC树脂：称取1.56g CTC Resin（如初始取代度约为0.64mmol/g）和1.2mmol Fmoc-Leu-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3.0mmol 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-Leu-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-Leu-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3.0mmol Fmoc-Lys(Boc)-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5.99mmol DIPEA，2.85mmol 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-Lys(Boc)-Leu-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-Lys(Boc)-Leu-CTC Resin

Fmoc-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

H2N-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

Fmoc-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

H2N-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

Fmoc-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

H2N-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

Fmoc-Ile-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

H2N-Ile-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

Fmoc-Ile-Ile-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

H2N-Ile-Ile-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-CTC Resin

Fmoc-Ser(tBu)-Ile-Ile-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-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-Ser(tBu)-Ile-Ile-Asn(Trt)-Phe-Glu(OtBu)-Lys(Boc)-Leu-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-Ser-Ile-Ile-Asn-Phe-Glu-Lys-Leu-COOH。结构图见产品结构图。

切割液选择：1）TFA：H2O=95%：5%、TFA：H2O=97.5%：2.5%

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

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