编号: | 145590 |
中文名称: | 胃泌素Big Gastrin 1 (human) |
英文名: | Big Gastrin 1 (human) |
CAS号: | 60675-77-6 |
单字母: | Pyr-LGPQGPPHLVADPSKKQGPWLEEEEEAYGWMDF-NH2 |
三字母: | Pyr 焦谷氨酸 -Leu亮氨酸 -Gly甘氨酸 -Pro脯氨酸 -Gln谷氨酰胺 -Gly甘氨酸 -Pro脯氨酸 -Pro脯氨酸 -His组氨酸 -Leu亮氨酸 -Val缬氨酸 -Ala丙氨酸 -Asp天冬氨酸 -Pro脯氨酸 -Ser丝氨酸 -Lys赖氨酸 -Lys赖氨酸 -Gln谷氨酰胺 -Gly甘氨酸 -Pro脯氨酸 -Trp色氨酸 -Leu亮氨酸 -Glu谷氨酸 -Glu谷氨酸 -Glu谷氨酸 -Glu谷氨酸 -Glu谷氨酸 -Ala丙氨酸 -Tyr酪氨酸 -Gly甘氨酸 -Trp色氨酸 -Met甲硫氨酸 -Asp天冬氨酸 -Phe苯丙氨酸 -NH2C端酰胺化 |
氨基酸个数: | 33 |
分子式: | C176H251N43O53S1 |
平均分子量: | 3849.2 |
精确分子量: | 3846.8 |
等电点(PI): | 4.17 |
pH=7.0时的净电荷数: | -3.76 |
平均亲水性: | 0.22083333333333 |
疏水性值: | -0.96 |
外观与性状: | 白色粉末状固体 |
消光系数: | 12490 |
来源: | 人工化学合成,仅限科学研究使用,不得用于人体。 |
纯度: | 95%、98% |
盐体系: | 可选TFA、HAc、HCl或其它 |
生成周期: | 2-3周 |
储存条件: | 负80℃至负20℃ |
标签: | 胃泌素 |
多肽Pyr-Leu-Gly-Pro-Gln-Gly-Pro-Pro-His-Leu-Val-Ala-Asp-Pro-Ser-Lys-Lys-Gln-Gly-Pro-Trp-Leu-Glu-Glu-Glu-Glu-Glu-Ala-Tyr-Gly-Trp-Met-Asp-Phe-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1.35g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1.22mmol 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2.43mmol DIPEA,1.15mmol 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-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-Phe-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Gly-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
H2N-Gly-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin
Fmoc-Leu-Gly-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-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-Leu-Gly-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-Linker-MBHA Resin,结构如下:
5、焦谷氨酸反应连接:在上述树脂中,加入适当DMF后,再加入1.22mmol 焦谷氨酸到树脂中,再加入2.43mmol DIPEA、1.15mmol 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Pyr-Leu-Gly-Pro-Gln(Trt)-Gly-Pro-Pro-His(Trt)-Leu-Val-Ala-Asp(OtBu)-Pro-Ser(tBu)-Lys(Boc)-Lys(Boc)-Gln(Trt)-Gly-Pro-Trp(Boc)-Leu-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Glu(OtBu)-Ala-Tyr(tBu)-Gly-Trp(Boc)-Met-Asp(OtBu)-Phe-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Pyr-Leu-Gly-Pro-Gln-Gly-Pro-Pro-His-Leu-Val-Ala-Asp-Pro-Ser-Lys-Lys-Gln-Gly-Pro-Trp-Leu-Glu-Glu-Glu-Glu-Glu-Ala-Tyr-Gly-Trp-Met-Asp-Phe-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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