作为 CCK 的拮抗剂，PP 抑制胰腺的沉淀。PP 还作用于胃肠蠕动、食物摄入和新陈代谢。
编号：158392
CAS号：1816258-36-2
单字母：Biotinyl-APLEPVYPGDNATPEQMAQYAADLRRYINMLTRPRY-CONH2
As antagonist of CCK, PP inhibits pancreatic sedretion. PP also acts on the gastrointestinal motility, food intake, and metabolism.
专肽生物合成用于蛋白质-蛋白质相互作用研究的生物素化肽。尽管生物素可以在 N 端或 C 端引入（通过赖氨酸残基），但我们建议使用 N 端修饰，因为它成本低、成功率高、周转时间短且易于操作。因为多肽合成是从 C 端到 N 端合成的，因此，N 端修饰是 SPPS步骤的最后一步，不需要额外的特定缩合步骤。相比之下，C 端修饰需要额外的步骤，并且通常更复杂。当然，原则上生物素可以定位在任何地方。
生物素可以通过多种不同的接头或间隔物与肽分离。尽管如此，还是建议包含一个灵活的间隔物，例如 Ahx（一个 6 碳接头），以使生物素标签更加稳定或灵活。
专肽生物在 N 端或 C 端提供生物素化：生物素-N 端、赖氨酸-生物素-肽中间和赖氨酸-生物素-C 端。
专肽生物还可以使用 Ahx 接头或长碳 (LC) 接头提供生物素化：生物素-Ahx-N 末端、Lys-Ahx-生物素-肽中间、Lys-Ahx-生物素-C-末端。
（生物素结构）
示例：
GRGDS在N端和C端标记生物素的结构展示。
1、GRGDS在N端标记生物素，不增加Ahx 接头
2、GRGDS在N端标记生物素，增加一个Ahx 接头
3、GRGDS在C端标记生物素，不增加Ahx 接头
4、GRGDS在C端标记生物素，增加一个Ahx 接头。
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| 10.1210/jc.2003-030630 | Pancreatic polypeptide reduces appetite and food intake in humans | 下载 |
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| 10.1038/oby.2006.180 | Physiological evidence for the involvement of peptide YY in the regulation of energy homeostasis in humans | 下载 |
| 10.1016/j.peptides.2012.06.008 | Surface masking shapes the traffic of the neuropeptide Y Y2 receptor | 下载 |
| 10.1002/oby.20534 | PYY3-36 and pancreatic polypeptide reduce food intake in an additive manner via distinct hypothalamic dependent pathways in mice | 下载 |
| 10.3390/ijms15034856 | Dimers of G-protein coupled receptors as versatile storage and response units | 下载 |
| 10.1021/acs.jmedchem.6b00310 | Optically Pure, Structural, and Fluorescent Analogues of a Dimeric Y4 Receptor Agonist Derived by an Olefin Metathesis Approach | 下载 |
| 10.1021/acs.bioconjchem.6b00376 | Identification of a Cyanine-Dye Labeled Peptidic Ligand for Y1R and Y4R, Based upon the Neuropeptide Y C-Terminal Analogue, BVD-15 | 下载 |
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多肽Biotin-Ala-Pro-Leu-Glu-Pro-Val-Tyr-Pro-Gly-Asp-Asn-Ala-Thr-Pro-Glu-Gln-Met-Ala-Gln-Tyr-Ala-Ala-Asp-Leu-Arg-Arg-Tyr-Ile-Asn-Met-Leu-Thr-Arg-Pro-Arg-Tyr-NH2的合成步骤：
1、合成MBHA树脂：取若干克MBHA树脂（如初始取代度为0.5mmol/g）和1倍树脂摩尔量的Fmoc-Linker-OH加入到反应器中，加入DMF，搅拌使氨基酸完全溶解。再加入树脂2倍量的DIEPA，搅拌混合均匀。再加入树脂0.95倍量的HBTU，搅拌混合均匀。反应3-4小时后，用DMF洗涤3次。用2倍树脂体积的10%乙酸酐/DMF 进行封端30分钟。然后再用DMF洗涤3次，甲醇洗涤2次，DCM洗涤2次，再用甲醇洗涤2次。真空干燥12小时以上，得到干燥的树脂{Fmoc-Linker-MHBA Resin}，测定取代度。这里测得取代度为 0.3mmol/g。结构如下图：
2、脱Fmoc：取2.1g的上述树脂，用DCM或DMF溶胀20分钟。用DMF洗涤2遍。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-Linker-MBHA 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1.89mmol Fmoc-Tyr(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3.78mmol DIPEA，1.8mmol 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-Tyr(tBu)-Linker-MBHA 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-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA 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-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin，结构如下：
5、生物素反应连接：在上述树脂中，加入适当DMF后，再加入1.89mmol 生物素到树脂中，再加入3.78mmol DIPEA、1.8mmol HBTU，鼓氮气反应30分钟。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Biotin-Ala-Pro-Leu-Glu(OtBu)-Pro-Val-Tyr(tBu)-Pro-Gly-Asp(OtBu)-Asn(Trt)-Ala-Thr(tBu)-Pro-Glu(OtBu)-Gln(Trt)-Met-Ala-Gln(Trt)-Tyr(tBu)-Ala-Ala-Asp(OtBu)-Leu-Arg(Pbf)-Arg(Pbf)-Tyr(tBu)-Ile-Asn(Trt)-Met-Leu-Thr(tBu)-Arg(Pbf)-Pro-Arg(Pbf)-Tyr(tBu)-Linker-MBHAResin。 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Biotin-Ala-Pro-Leu-Glu-Pro-Val-Tyr-Pro-Gly-Asp-Asn-Ala-Thr-Pro-Glu-Gln-Met-Ala-Gln-Tyr-Ala-Ala-Asp-Leu-Arg-Arg-Tyr-Ile-Asn-Met-Leu-Thr-Arg-Pro-Arg-Tyr-NH2。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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