浙江省科技型企業(yè)---加速您的多肽研究
400-998-5282
專注多肽 服務(wù)科研
400-998-5282
專注多肽 服務(wù)科研
編號(hào)：134106
CAS號(hào)：178861-83-1
單字母：H2N-YPSKPDNPGEDAPAEDMARYYSALRHYINLIwRQRY-CONH2
很多蛋白在細(xì)胞中非常容易被降解，或被標(biāo)記，進(jìn)而被選擇性地破壞。但含有部分D型氨基酸的多肽則顯示了很強(qiáng)的抵抗蛋白酶降解能力。
多肽H2N-Tyr-Pro-Ser-Lys-Pro-Asp-Asn-Pro-Gly-Glu-Asp-Ala-Pro-Ala-Glu-Asp-Met-Ala-Arg-Tyr-Tyr-Ser-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-DTrp-Arg-Gln-Arg-Tyr-NH2的合成步驟：
1、合成MBHA樹脂：取若干克的MBHA樹脂（如初始取代度為0.5mmol/g）和1倍樹脂摩爾量的Fmoc-Linker-OH加入到反應(yīng)器中，加入DMF，攪拌使氨基酸完全溶解。再加入樹脂2倍量的DIEPA，攪拌混合均勻。再加入樹脂0.95倍量的HBTU，攪拌混合均勻。反應(yīng)3-4小時(shí)后，用DMF洗滌3次。用2倍樹脂體積的10%乙酸酐/DMF 進(jìn)行封端30分鐘。然后再用DMF洗滌3次，甲醇洗滌2次，DCM洗滌2次，再用甲醇洗滌2次。真空干燥12小時(shí)以上，得到干燥的樹脂{Fmoc-Linker-MHBA Resin}，測定取代度。這里測得取代度為 0.3mmol/g。結(jié)構(gòu)如下圖：
2、脫Fmoc：取2.14g的上述樹脂，用DCM或DMF溶脹20分鐘。用DMF洗滌2遍。加3倍樹脂體積的20%Pip/DMF溶液，鼓氮?dú)?0分鐘，然后2倍樹脂體積的DMF 洗滌5次。得到 H2N-Linker-MBHA Resin 。（此步驟脫除Fmoc基團(tuán)，茚三酮檢測為藍(lán)色，Pip為哌啶）。結(jié)構(gòu)圖如下：
3、縮合：取1.93mmol Fmoc-Tyr(tBu)-OH 氨基酸，加入到上述樹脂里，加適當(dāng)DMF溶解氨基酸，再依次加入3.85mmol DIPEA，1.83mmol HBTU。反應(yīng)30分鐘后，取小樣洗滌，茚三酮檢測為無色。用2倍樹脂體積的DMF 洗滌3次樹脂。（洗滌樹脂，去掉殘留溶劑，為下一步反應(yīng)做準(zhǔn)備）。得到Fmoc-Tyr(tBu)-Linker-MBHA Resin。氨基酸：DIPEA：HBTU：樹脂=3：6：2.85：1（摩爾比）。結(jié)構(gòu)圖如下：
4、依次循環(huán)步驟二、步驟三，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-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Ser(tBu)-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Ser(tBu)-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Pro-Ser(tBu)-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
H2N-Pro-Ser(tBu)-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Pro-Ser(tBu)-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin
以上中間結(jié)構(gòu)，均可在專肽生物多肽計(jì)算器-多肽結(jié)構(gòu)計(jì)算器中，一鍵畫出。
最后再經(jīng)過步驟二得到 H2N-Tyr(tBu)-Pro-Ser(tBu)-Lys(Boc)-Pro-Asp(OtBu)-Asn(Trt)-Pro-Gly-Glu(OtBu)-Asp(OtBu)-Ala-Pro-Ala-Glu(OtBu)-Asp(OtBu)-Met-Ala-Arg(Pbf)-Tyr(tBu)-Tyr(tBu)-Ser(tBu)-Ala-Leu-Arg(Pbf)-His(Trt)-Tyr(tBu)-Ile-Asn(Trt)-Leu-Ile-DTrp(Boc)-Arg(Pbf)-Gln(Trt)-Arg(Pbf)-Tyr(tBu)-Linker-MBHA Resin，結(jié)構(gòu)如下：
5、切割：6倍樹脂體積的切割液（或每1g樹脂加8ml左右的切割液），搖床搖晃 2小時(shí)，過濾掉樹脂，用冰無水乙醚沉淀濾液，并用冰無水乙醚洗滌沉淀物3次，最后將沉淀物放真空干燥釜中，常溫干燥24小試，得到粗品H2N-Tyr-Pro-Ser-Lys-Pro-Asp-Asn-Pro-Gly-Glu-Asp-Ala-Pro-Ala-Glu-Asp-Met-Ala-Arg-Tyr-Tyr-Ser-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-DTrp-Arg-Gln-Arg-Tyr-NH2。結(jié)構(gòu)圖見產(chǎn)品結(jié)構(gòu)圖。
切割液選擇：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%
（前兩種適合沒有容易氧化的氨基酸，例如Trp、Cys、Met。第三種適合幾乎所有的序列。）
6、純化凍干：使用液相色譜純化，收集目標(biāo)峰液體，進(jìn)行凍干，獲得蓬松的粉末狀固體多肽。不過這時(shí)要取小樣復(fù)測下純度 是否目標(biāo)純度。
7、最后總結(jié)：
杭州專肽生物技術(shù)有限公司（ALLPEPTIDE https://www.allpeptide.com）主營定制多肽合成業(yè)務(wù)，提供各類長肽，短肽，環(huán)肽，提供各類修飾肽，如：熒光標(biāo)記修飾（CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等），功能基團(tuán)修飾肽（疊氮、炔基、DBCO、DOTA、NOTA等），同位素標(biāo)記肽（N15、C13），訂書肽（Stapled Peptide）,脂肪酸修飾肽（Pal、Myr、Ste），磷酸化修飾肽（P-Ser、P-Thr、P-Tyr），環(huán)肽（酰胺鍵環(huán)肽、一對(duì)或者多對(duì)二硫鍵環(huán)），生物素標(biāo)記肽，PEG修飾肽，甲基化修飾肽等。
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![神經(jīng)肽Y [DTrp32]Neuro Peptide Y(1-36), human,H2N-Tyr-Pro-Ser-Lys-Pro-Asp-Asn-Pro-Gly-Glu-Asp-Ala-Pro-Ala-Glu-Asp-Met-Ala-Arg-Tyr-Tyr-Ser-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu-Ile-DTrp-Arg-Gln-Arg-Tyr-NH2,H2N-YPSKPDNPGEDAPAEDMARYYSALRHYINLI-DTrp-RQRY-NH2,安徽專肽生物的產(chǎn)品](/static/ah/images/def.jpg)
