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1802086-20-9,淀粉肽Biotin-β-Amyloid(1-42)、,Biotin-Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-COOH,Biotin-DAEFRHDSGYEVHHQKLVFFAEDVGSNK,杭州專肽生物的產(chǎn)品

淀粉肽Biotin-β-Amyloid(1-42)、

N-末端生物素標(biāo)記的 Aβ42。

編號(hào):163583

CAS號(hào):1802086-20-9

單字母:Biotinyl-DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA-OH

糾錯(cuò)
  • 編號(hào):163583
    中文名稱:淀粉肽Biotin-β-Amyloid(1-42)、
    英文名:Biotin-β-Amyloid(1-42)
    CAS號(hào):1802086-20-9
    單字母:Biotinyl-DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA-OH
    三字母:Biotinyl

    N端生物素標(biāo)記

    -Asp

    天冬氨酸

    -Ala

    丙氨酸

    -Glu

    谷氨酸

    -Phe

    苯丙氨酸

    -Arg

    精氨酸

    -His

    組氨酸

    -Asp

    天冬氨酸

    -Ser

    絲氨酸

    -Gly

    甘氨酸

    -Tyr

    酪氨酸

    -Glu

    谷氨酸

    -Val

    纈氨酸

    -His

    組氨酸

    -His

    組氨酸

    -Gln

    谷氨酰胺

    -Lys

    賴氨酸

    -Leu

    亮氨酸

    -Val

    纈氨酸

    -Phe

    苯丙氨酸

    -Phe

    苯丙氨酸

    -Ala

    丙氨酸

    -Glu

    谷氨酸

    -Asp

    天冬氨酸

    -Val

    纈氨酸

    -Gly

    甘氨酸

    -Ser

    絲氨酸

    -Asn

    天冬酰胺

    -Lys

    賴氨酸

    -Gly

    甘氨酸

    -Ala

    丙氨酸

    -Ile

    異亮氨酸

    -Ile

    異亮氨酸

    -Gly

    甘氨酸

    -Leu

    亮氨酸

    -Met

    甲硫氨酸

    -Val

    纈氨酸

    -Gly

    甘氨酸

    -Gly

    甘氨酸

    -Val

    纈氨酸

    -Val

    纈氨酸

    -Ile

    異亮氨酸

    -Ala

    丙氨酸

    -OH

    C端羧基

    氨基酸個(gè)數(shù):42
    分子式:C213H325N57O62S2
    平均分子量:4740.33
    精確分子量:4737.35
    等電點(diǎn)(PI):6.23
    pH=7.0時(shí)的凈電荷數(shù):-1.29
    平均親水性:-0.15833333333333
    疏水性值:0.21
    外觀與性狀:白色粉末狀固體
    消光系數(shù):1490
    來(lái)源:人工化學(xué)合成,僅限科學(xué)研究使用,不得用于人體。
    純度:95%、98%
    鹽體系:可選TFA、HAc、HCl或其它
    生成周期:2-3周
    儲(chǔ)存條件:負(fù)80℃至負(fù)20℃
    標(biāo)簽:生物素標(biāo)記肽(Biotinyl)    淀粉樣肽(Amyloid Peptides)   

  • N-terminally biotin-labeled Aβ42.

    專肽生物合成用于蛋白質(zhì)-蛋白質(zhì)相互作用研究的生物素化肽。盡管生物素可以在 N 端或 C 端引入(通過(guò)賴氨酸殘基),但我們建議使用 N 端修飾,因?yàn)樗杀镜?、成功率高、周轉(zhuǎn)時(shí)間短且易于操作。因?yàn)槎嚯暮铣墒菑?C 端到 N 端合成的,因此,N 端修飾是 SPPS步驟的最后一步,不需要額外的特定縮合步驟。相比之下,C 端修飾需要額外的步驟,并且通常更復(fù)雜。當(dāng)然,原則上生物素可以定位在任何地方

    生物素可以通過(guò)多種不同的接頭或間隔物與肽分離。盡管如此,還是建議包含一個(gè)靈活的間隔物,例如 Ahx(一個(gè) 6 碳接頭),以使生物素標(biāo)簽更加穩(wěn)定或靈活。

    專肽生物在 N 端或 C 端提供生物素化:生物素-N 端、賴氨酸-生物素-肽中間和賴氨酸-生物素-C 端。
    專肽生物還可以使用 Ahx 接頭或長(zhǎng)碳 (LC) 接頭提供生物素化:生物素-Ahx-N 末端、Lys-Ahx-生物素-肽中間、Lys-Ahx-生物素-C-末端。

    (生物素結(jié)構(gòu))

    示例:
    GRGDS在N端和C端標(biāo)記生物素的結(jié)構(gòu)展示。

    1、GRGDS在N端標(biāo)記生物素,不增加Ahx 接頭

    2、GRGDS在N端標(biāo)記生物素,增加一個(gè)Ahx 接頭


    3、GRGDS在C端標(biāo)記生物素,不增加Ahx 接頭

    4、GRGDS在C端標(biāo)記生物素,增加一個(gè)Ahx 接頭。

    淀粉肽背景:β淀粉樣蛋白(Aβ或Abeta)是從淀粉樣前體蛋白加工而成的含有36–43個(gè)氨基酸的多肽。Aβ是與阿爾茲海默病相關(guān)的淀粉樣蛋白斑的成分。已有證據(jù)表明,Aβ是一個(gè)多功能肽,具有顯著的非病理性活性。Aβ是阿爾茲海默病患者腦中發(fā)現(xiàn)的沉積物的主要成分。在散發(fā)性阿爾茲海默病患者的腦中,Aβ的水平升高,造成腦血管病變和神經(jīng)毒性。Aβ蛋白是由β和γ分泌酶的連續(xù)作用而產(chǎn)生的。γ分泌酶產(chǎn)生Aβ肽的C末端,在APP的轉(zhuǎn)膜結(jié)構(gòu)域切割,可以產(chǎn)生許多36-43個(gè)氨基酸殘基長(zhǎng)度的異構(gòu)體,最常見(jiàn)的異構(gòu)體是Aβ40和Aβ42。更長(zhǎng)形式的Aβ在內(nèi)質(zhì)網(wǎng)中切割產(chǎn)生,而更短形式的Aβ在反面高爾基網(wǎng)中產(chǎn)生。

    structure of Amyloid β-Peptide (1-40) (human)

    淀粉樣蛋白肽的 定義淀粉樣蛋白 是絲狀蛋白質(zhì)沉積物,大小從納米到微米不等,并且由肽β鏈的平行或反平行排列形成的聚集的肽β折疊構(gòu)成。

    結(jié)構(gòu)特征:使用固態(tài)NMR(SSNMR),與計(jì)算能量最小化過(guò)程結(jié)合,Tycko和合作者已經(jīng)提出從淀粉狀蛋白肽SS(Aß1-40)的40個(gè)殘基的形式形成的淀粉樣蛋白原纖維的結(jié)構(gòu)在pH 7.4和24 o C在靜止條件下。在這種結(jié)構(gòu)中,每個(gè)Aß1-40分子在原纖維的核心區(qū)域貢獻(xiàn)一對(duì)ß鏈,大約跨越殘基12-24和30-40。這些由回路25-29連接的鏈不是同一張ß-sheet的一部分,但參與同一原絲內(nèi)兩個(gè)不同的ß-sheets的形成。不同的Aß分子2、3至少?gòu)牡?到39位殘基以平行排列和對(duì)齊的方式相互堆疊。通過(guò)調(diào)用其他實(shí)驗(yàn)約束,例如使用透射電子顯微鏡(TEM)觀察到的原絲直徑和單位質(zhì)量通過(guò)掃描透射電子顯微鏡(STEM)1、2測(cè)得的長(zhǎng)度表明,單個(gè)原絲是由四個(gè)ß片組成的,它們之間的距離約為10Å。

    作用模式:阿爾茨海默氏病(AD)是淀粉樣蛋白絲狀沉積物的結(jié)果,淀粉狀蛋白沉積物在分子水平上定義該疾病,發(fā)生在神經(jīng)周膜,軸突,樹(shù)突和神經(jīng)元末端,如神經(jīng)原纖維纏結(jié)(NFT),在細(xì)胞外神經(jīng)纖維中淀粉樣斑塊(APC),以及周圍的血管稱為淀粉樣嗜血性血管?。ˋCA)。淀粉樣蛋白沉積物顯然發(fā)生在發(fā)展NFT的神經(jīng)元末端區(qū)域。已經(jīng)表明,APC和ACA的主要成分已被證明是4.5kDa的淀粉樣蛋白,最初被稱為“β-蛋白”或“淀粉樣蛋白A4”,我們現(xiàn)在將其稱為“βA4”。

    功能:鈣失調(diào)和膜破壞是可溶性淀粉樣蛋白低聚物普遍存在的神經(jīng)毒性機(jī)制:進(jìn)行了一項(xiàng)研究,以研究Ca 2+信號(hào)轉(zhuǎn)導(dǎo)可能參與淀粉樣蛋白誘導(dǎo)的細(xì)胞毒性,疾病相關(guān)淀粉樣蛋白(β,病毒,胰島淀粉樣蛋白)的均質(zhì)制劑制備了處于各種聚集狀態(tài)的多肽,聚谷氨酰胺和溶菌酶),并測(cè)試了它們對(duì)加載fluo-3的SH-SY5Y細(xì)胞的作用。寡聚形式的所有淀粉樣蛋白的應(yīng)用(0.6-6 µg / ml)迅速(約5 s)使細(xì)胞內(nèi)Ca 2+升高,而等量的單體和原纖維則沒(méi)有。細(xì)胞內(nèi)Ca 2+耗盡后,Abeta42低聚物引起的Ca 2+信號(hào)持續(xù)存在店,和小信號(hào)仍留在鈣2 + -游離介質(zhì),指示從細(xì)胞外和細(xì)胞內(nèi)Ca貢獻(xiàn)2+源。膜對(duì)Ca 2+的滲透性增加不能歸因于內(nèi)源性Ca 2+通道的活化,因?yàn)榉磻?yīng)不受強(qiáng)力的Ca 2 +-通道阻滯劑鈷的影響。取而代之的是,觀察到Abeta42和其他低聚物引起陰離子熒光染料的快速細(xì)胞泄漏,這表明膜通透性普遍提高。導(dǎo)致的離子和分子通量失調(diào)可能為許多淀粉樣變性疾病中Ca 2+失調(diào)提供了由低聚物介導(dǎo)的毒性的常見(jiàn)機(jī)制。離子起著至關(guān)重要的作用,因?yàn)樗鼈兊目缒舛忍荻群軓?qiáng),并且參與了細(xì)胞功能障礙和死亡。

    2型糖尿病中的胰島淀粉樣蛋白和毒性低聚物假說(shuō): 2型糖尿?。═2DM)的特征是胰島素抵抗,胰島素分泌缺陷,β細(xì)胞量減少,β細(xì)胞凋亡增加和胰島淀粉樣蛋白。胰島淀粉樣蛋白源自胰島淀粉樣蛋白多肽(IAPP,胰島淀粉樣多肽),該蛋白是通過(guò)胰β細(xì)胞與胰島素共表達(dá)和共分泌的蛋白。與其他淀粉樣蛋白一樣,IAPP具有形成膜滲透性毒性低聚物的傾向。越來(lái)越多的證據(jù)表明,這些有毒的寡聚體而不是這些蛋白質(zhì)的細(xì)胞外淀粉樣蛋白形式,是導(dǎo)致神經(jīng)退行性疾病中神經(jīng)元丟失的原因。有人提出,胞內(nèi)IAPP寡聚物的形成可能會(huì)導(dǎo)致T2DM 6中的β細(xì)胞丟失。

  • DOI名稱
    10.1093/brain/awh531LPS receptor (CD14): a receptor for phagocytosis of Alzheimer's amyloid peptide下載
    10.1073/pnas.0703137104Liposomal vaccines with conformation-specific amyloid peptide antigens define immune response and efficacy in APP transgenic mice下載
    10.1186/1750-1326-3-1Ginkgolides protect against amyloid-beta1-42-mediated synapse damage in vitro下載
    10.1002/anie.200904902Identification of hot regions of the Abeta-IAPP interaction interface as high-affinity binding sites in both cross- and self-association下載
    10.3233/JAD-2012-120535GABA attenuates amyloid toxicity by downregulating its endocytosis and improves cognitive impairment下載
    10.1021/cn400146aBenzothiazole aniline tetra(ethylene glycol) and 3-amino-1,2,4-triazole inhibit neuroprotection against amyloid peptides by catalase overexpression in vitro下載
    10.1016/j.biomaterials.2013.11.028A 3D Alzheimer's disease culture model and the induction of P21-activated kinase mediated sensing in iPSC derived neurons下載
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  • 多肽Biotin-Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-COOH的合成步驟:

    1、合成CTC樹(shù)脂:稱取1.33g CTC Resin(如初始取代度約為0.4mmol/g)和0.64mmol Fmoc-Ala-OH于反應(yīng)器中,加入適量DCM溶解氨基酸(需要注意,此時(shí)CTC樹(shù)脂體積會(huì)增大好幾倍,避免DCM溶液過(guò)少),再加入1.6mmol DIPEA(Mw:129.1,d:0.740g/ml),反應(yīng)2-3小時(shí)后,可不抽濾溶液,直接加入1ml的HPLC級(jí)甲醇,封端半小時(shí)。依次用DMF洗滌2次,甲醇洗滌1次,DCM洗滌一次,甲醇洗滌一次,DCM洗滌一次,DMF洗滌2次(這里使用甲醇和DCM交替洗滌,是為了更好地去除其他溶質(zhì),有利于后續(xù)反應(yīng))。得到  Fmoc-Ala-CTC Resin。結(jié)構(gòu)圖如下:

    2、脫Fmoc:加3倍樹(shù)脂體積的20%Pip/DMF溶液,鼓氮?dú)?0分鐘,然后2倍樹(shù)脂體積的DMF 洗滌5次。得到 H2N-Ala-CTC Resin 。(此步驟脫除Fmoc基團(tuán),茚三酮檢測(cè)為藍(lán)色,Pip為哌啶)。結(jié)構(gòu)圖如下:

    3、縮合:取1.6mmol Fmoc-Ile-OH 氨基酸,加入到上述樹(shù)脂里,加適當(dāng)DMF溶解氨基酸,再依次加入3.19mmol DIPEA,1.52mmol HBTU。反應(yīng)30分鐘后,取小樣洗滌,茚三酮檢測(cè)為無(wú)色。用2倍樹(shù)脂體積的DMF 洗滌3次樹(shù)脂。(洗滌樹(shù)脂,去掉殘留溶劑,為下一步反應(yīng)做準(zhǔn)備)。得到Fmoc-Ile-Ala-CTC Resin。氨基酸:DIPEA:HBTU:樹(shù)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-Ile-Ala-CTC Resin

    Fmoc-Val-Ile-Ala-CTC Resin

    H2N-Val-Ile-Ala-CTC Resin

    Fmoc-Val-Val-Ile-Ala-CTC Resin

    H2N-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Ala-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    H2N-Ala-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin

    Fmoc-Asp(OtBu)-Ala-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC 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(guò)步驟二得到 H2N-Asp(OtBu)-Ala-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTC Resin,結(jié)構(gòu)如下:

    5、生物素反應(yīng)連接:在上述樹(shù)脂中,加入適當(dāng)DMF后,再加入1.6mmol 生物素到樹(shù)脂中,再加入3.19mmol DIPEA、1.52mmol HBTU,鼓氮?dú)夥磻?yīng)30分鐘。用2倍樹(shù)脂體積的DMF 洗滌3次樹(shù)脂(洗滌樹(shù)脂,去掉殘留溶劑,為下一步反應(yīng)做準(zhǔn)備)。 得到Biotin-Asp(OtBu)-Ala-Glu(OtBu)-Phe-Arg(Pbf)-His(Trt)-Asp(OtBu)-Ser(tBu)-Gly-Tyr(tBu)-Glu(OtBu)-Val-His(Trt)-His(Trt)-Gln(Trt)-Lys(Boc)-Leu-Val-Phe-Phe-Ala-Glu(OtBu)-Asp(OtBu)-Val-Gly-Ser(tBu)-Asn(Trt)-Lys(Boc)-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-CTCResin。 結(jié)構(gòu)如下:

    6、切割:6倍樹(shù)脂體積的切割液(或每1g樹(shù)脂加8ml左右的切割液),搖床搖晃 2小時(shí),過(guò)濾掉樹(shù)脂,用冰無(wú)水乙醚沉淀濾液,并用冰無(wú)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Biotin-Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-Ile-Ala-COOH。結(jié)構(gòu)圖見(jiàn)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(méi)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(guò)這時(shí)要取小樣復(fù)測(cè)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 http://amynixphotography.com)主營(yíng)定制多肽合成業(yè)務(wù),提供各類長(zhǎng)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(shū)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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