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MAGE-3 (271-279) 是一个由 MAGE-3 编码的黑色素瘤抗原的 271-279 序列的短肽。MAGE-3 是一种 CTL 定义的 MAGE-3 蛋白，与人类白细胞抗原 (HLA)-A2 分子相关。MAGE-3 在包括恶性黑色素瘤在内的不同人类肿瘤类型中过表达，但在睾丸和胎盘以外的正常组织中没有过表达。
MAGE-3 (271-279) is a 271-279 residue peptide derived from melanoma antigens encoded by MAGE-3. MAGE-3 is a cytolytic T lymphocyte (CTL)-defined MAGE-3 protein associated with the human leukocyte antigen (HLA)-A2 molecule. MAGE-3 is overexpressed in different human tumor types, including malignant melanoma, but not by normal tissues except for testis and placenta[1].

ALPHA FACTOR SIGNALING PEPTIDE\n\n\nYeast mating signaling pathway\n\n\n\nThe life cycle of Saccharomyces cerevisiae, also known as the brewer’s and the baker’s yeast, is characterized by both haploid and diploid phases. Indeed, yeasts are organized in two haploid types, known as a and α cells (MATa and MATα), that can either proliferate individually, using mitosis, or fuse together to form MATa/MATα diploids.\n\nα factor - WHWLQLKPGQPMY peptide - CAS: 59401-28-4. Haploid, diploid cells\n\nFor two haploid cells to mate, mating factors are needed. These pheromones induce G-Protein Coupled Receptor‘s (GPCR) conformational changes, which allow G-protein migration in the plasma membrane. GPCRs are present on both a and α yeast cells and turn into Guanine nucleotide Exchange Factors (GEF) upon G-protein release. The receptor’s « new activity » leads to guanosine diphosphate (GDP) exchange by guanosine triphosphate (GTP) on the G-protein, which will recruit STE 5 to allow Fus3 protein phosphorylation. Phosphate addition on Fus3 enables its fusion to the cell’s plasma membrane in order to activate formin proteins by phosphorylation.\n\nα factor - WHWLQLKPGQPMY peptide - CAS: 59401-28-4. Yeast mating signaling pathway\n\nFormin proteins being involved in the polymerization of microfilaments, their consequent growth within the cell will form shmoo projections to enable complete mating of MATa and MATα cells, towards MATa/MATα diploid cell that can undergo mitosis and meiosis to create new haploid spores.\n\nα factor - WHWLQLKPGQPMY - CAS: 59401-28-4. Yeast mating\n\n\nCAS: 59401-28-4 α-factor pheromone – WHWLQLKPGQPMY\n\n\n\nTo induce a yeast cells’ shmoo projection, WHWLQLKPGQPMY pheromone (or alpha mating factor) is produced by α yeast cells. This signaling peptide is part of the MFAL1_YEAST protein (UniProt: P01149) that excretes its active factor into the culture medium to act on the opposite mating type).\n\nα mating factor is a 13 residues peptide inducing the expression of necessary mating genes ( 200 genes – 3% of the yeast’s genome) to arrest yeast cell cycle in the G1 phase, while altering cell surface and nuclear determinants, towards MATa and MATα fusion.\n\nWHWLQLKPGQPMY interacts with its complementary pheromone, a-factor, which is a 12 residues peptide (YIIKGVFWDPAC) that is covalently attached to farnesyl to generate the appropriate response upon α pheromone activation.\n\n\nAlpha factor applications\n\n\n\nSB-PEPTIDE offers α mating factor WHWLQLKPGQPMY to study:\n\n\n\n Cell cycle\n Cellular morphology\n Transcriptional induction\n Signal transduction pathways

J Herman, et al. A peptide encoded by the human MAGE3 gene and presented by HLA-B44 induces cytolytic T lymphocytes that recognize tumor cells expressing MAGE3. Immunogenetics. 1996;43(6):377-83. : https://pubmed.ncbi.nlm.nih.gov/8606058/

多肽H2N-Phe-Leu-Trp-Gly-Pro-Arg-Ala-Leu-Val-COOH的合成步骤：

1、合成CTC树脂：称取1.59g CTC Resin（如初始取代度约为1.02mmol/g）和1.95mmol Fmoc-Val-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4.87mmol 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-Val-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-Val-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4.87mmol Fmoc-Leu-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9.73mmol DIPEA，4.62mmol 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-Leu-Val-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-Leu-Val-CTC Resin

Fmoc-Ala-Leu-Val-CTC Resin

H2N-Ala-Leu-Val-CTC Resin

Fmoc-Arg(Pbf)-Ala-Leu-Val-CTC Resin

H2N-Arg(Pbf)-Ala-Leu-Val-CTC Resin

Fmoc-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

H2N-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

Fmoc-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

H2N-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

Fmoc-Trp(Boc)-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

H2N-Trp(Boc)-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

Fmoc-Leu-Trp(Boc)-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

H2N-Leu-Trp(Boc)-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-CTC Resin

Fmoc-Phe-Leu-Trp(Boc)-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-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-Leu-Trp(Boc)-Gly-Pro-Arg(Pbf)-Ala-Leu-Val-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-Leu-Trp-Gly-Pro-Arg-Ala-Leu-Val-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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