400-998-5282
专注多肽 服务科研
400-998-5282
专注多肽 服务科研
淋巴细胞性脉络丛脑膜炎病毒 (LCMV) 的 396 至 404 片段。
编号：200509
CAS号：158475-79-7
单字母：H2N-FQPQNGQFI-OH
Nucleoprotein (396-404) 是淋巴细胞性脉络丛脑膜炎病毒 (LCMV) 的 396 至 404 片段。 Nucleoprotein (396-404) 是 H-2D(b) 限制性免疫显性表位，可用作病毒抗原的分子模型。
Nucleoprotein (396-404) is the 396 to 404 fragment of lymphocytic choriomeningitis virus (LCMV). Nucleoprotein (396-404) is the H-2D(b)-restricted immunodominant epitope and can be used as a molecular model of viral antigen[1][2] .
Peptide H-FQPQNGQFI-OH is a Research Peptide with significant interest within the field academic and medical research. Recent citations using H-FQPQNGQFI-OH include the following: A cell-based MHC stabilization assay for the detection of peptide binding to the canine classical class I molecule, DLA-88 P Ross, JC Holmes, GS Gojanovich, PR Hess - Veterinary immunology and , 2012 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0165242712003340 Thymic selection and adaptability of cytotoxic T lymphocyte responses in transgenic mice expressing a viral protein in the thymus. MG Von Herrath, J Dockter, M Nerenberg - The Journal of , 1994 - rupress.orghttps://rupress.org/jem/article-abstract/180/5/1901/25509 Use of a high-affinity peptide that aborts MHC-restricted cytotoxic T lymphocyte activity against multiple viruses in vitro and virus-induced immunopathologic disease in MBA Oldstone, M von Herrath, H Lewicki, D Hudrisier - Virology, 1999 - Elsevierhttps://www.sciencedirect.com/science/article/pii/S0042682298995939 Molecular Anatomy of Autoimmune Disease MBA Oldstone - Symposium in Immunology V: Antiviral Immunity, 1996 - Springerhttps://link.springer.com/chapter/10.1007/978-3-642-79896-2_2 Discriminated selection among viral peptides with the appropriate anchor residues: implications for the size of the cytotoxic T-lymphocyte repertoire and control of viral MB Oldstone, H Lewicki, P Borrow, D Hudrisier - Journal of , 1995 - Am Soc Microbiolhttps://journals.asm.org/doi/abs/10.1128/jvi.69.12.7423-7429.1995 Optimal lymphocytic choriomeningitis virus sequences restricted by H-2Db major histocompatibility complex class I molecules and presented to cytotoxic T JE Gairin, H Mazarguil, D Hudrisier - Journal of , 1995 - Am Soc Microbiolhttps://journals.asm.org/doi/abs/10.1128/jvi.69.4.2297-2305.1995 Thymic Selection and Adaptability of Cytotoxic T Lymphocyte Responses in Transgenic Mice Expressing a Viral Protein in the Thymus By Matthias G. yon Herrath JE Gairin - ncbi.nlm.nih.govhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2191719/ T cells infiltrate the brain in murine and human transmissible spongiform encephalopathies H Lewicki, A Tishon, D Homann , H Mazarguil - Journal of , 2003 - Am Soc Microbiolhttps://journals.asm.org/doi/abs/10.1128/jvi.77.6.3799-3808.2003 Tumor necrosis factor receptor p55 mediates deletion of peripheral cytotoxic T lymphocytes in vivo DE Speiser , E Sebzda, T Ohteki - European journal of , 1996 - Wiley Online Libraryhttps://onlinelibrary.wiley.com/doi/abs/10.1002/eji.1830261235 Structural and functional identification of major histocompatibility complex class I-restricted self-peptides as naturally occurring molecular mimics of viral antigens D Hudrisier , J Riond , O Burlet-Schiltz - Journal of Biological , 2001 - ASBMBhttps://www.jbc.org/article/S0021-9258(19)67082-1/abstract Evidence for an underlying CD4 helper and CD8 T-cell defect in B-cell-deficient mice: failure to clear persistent virus infection after adoptive immunotherapy with virus D Homann , A Tishon, DP Berger, WO Weigle - Journal of , 1998 - Am Soc Microbiolhttps://journals.asm.org/doi/abs/10.1128/jvi.72.11.9208-9216.1998 Puromycin-sensitive aminopeptidase limits MHC class I presentation in dendritic cells but does not affect CD8 T cell responses during viral infections CF Towne, IA York , J Neijssen , ML Karow - The Journal of , 2008 - journals.aai.orghttps://journals.aai.org/jimmunol/article/180/3/1704/75825 Major differences in transporter associated with antigen presentation (TAP)-dependent translocation of MHC class I-presentable peptides and the effect of flanking A Neisig, J Roelse, AJ Sijts , F Ossendorp - (Baltimore, Md.: 1950 , 1995 - journals.aai.orghttps://journals.aai.org/jimmunol/article-abstract/154/3/1273/5936 Protective immunity does not correlate with the hierarchy of virus-specific cytotoxic T cell responses to naturally processed peptides A Gallimore, T Dumrese, H Hengartner - The Journal of , 1998 - rupress.orghttps://rupress.org/jem/article-abstract/187/10/1647-b/7353
| DOI | 名称 | |
|---|---|---|
| 10.1074/jbc.M008864200 | Structural and functional identification of major histocompatibility complex class I-restricted self-peptides as naturally occurring molecular mimics of viral antigens. Possible role in CD8+ T cell-mediated, virus-induced autoimmune disease | 下载 |
| 10.1128/JVI.69.4.2297-2305.1995 | Optimal lymphocytic choriomeningitis virus sequences restricted by H-2Db major histocompatibility complex class I molecules and presented to cytotoxic T lymphocytes | 下载 |
多肽H2N-Phe-Gln-Pro-Gln-Asn-Gly-Gln-Phe-Ile-COOH的合成步骤：
1、合成CTC树脂：称取2.7g CTC Resin（如初始取代度约为0.52mmol/g）和1.68mmol Fmoc-Ile-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4.21mmol 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-Ile-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-Ile-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4.21mmol Fmoc-Phe-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8.42mmol DIPEA，4.0mmol 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-Phe-Ile-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-Phe-Ile-CTC Resin
Fmoc-Gln(Trt)-Phe-Ile-CTC Resin
H2N-Gln(Trt)-Phe-Ile-CTC Resin
Fmoc-Gly-Gln(Trt)-Phe-Ile-CTC Resin
H2N-Gly-Gln(Trt)-Phe-Ile-CTC Resin
Fmoc-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
H2N-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
Fmoc-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
H2N-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
Fmoc-Pro-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
H2N-Pro-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
Fmoc-Gln(Trt)-Pro-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
H2N-Gln(Trt)-Pro-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-CTC Resin
Fmoc-Phe-Gln(Trt)-Pro-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-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-Phe-Gln(Trt)-Pro-Gln(Trt)-Asn(Trt)-Gly-Gln(Trt)-Phe-Ile-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-Phe-Gln-Pro-Gln-Asn-Gly-Gln-Phe-Ile-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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