专肽生物_定制多肽合成服务_多肽合成定制服务_多肽合成厂家_多肽合成公司

KRSR selectively enhanced heparan sulfate-mediated osteoblast adhesion mechanisms. Elucidating such peptide sequence is of importance to the field of bone-cell/tissue engineering as well as to the design of proactive dental/orthopedic biomaterials. Note: Uses of this peptide are covered by U.S. Patent No. 6,262,017 B1 owned by Rensselaer Polytechnic Institute (RPI) located in Troy, New York, U.S.A. RPI authorizes purchasers of 5 grams or less per year to use this peptide for research purposes only. Other purchasers must have a written licence from RPI.

Laminins are the protein network that is the foundation for most cells and organs. They are large trimeric proteins, also in the family of glycoproteins that contain an alpha chain, a beta chain and a gamma chain which are found in five, three and three genetic variants. As a family of glycoproteins, laminins are an integral part of the structural scaffolding in almost every tissue of an organism. This makes the laminin a vital part of the maintenance and survival of the tissues. Consequently, defective laminins cause muscular dystrophy, junctional epidermolysis bullosa and defects of the kidney a defect.

Recently, publications have shown that laminins can be used to culture cells that are difficult to culture on other substrates. These trimeric glycoproteins form a similar structure to a cross, giving it a structure that has the ability to bind to other cell membrane and extracellular molecules. The three shorter arms of the laminin are particularly efficient at binding to other laminin molecules, which allows them to form larger sheets. The long arm is capable of binding to cells, which then helps anchor organized tissue cells to the membrane. These laminins are an important and biologically active part of the basal lamina, influenincing cell differentiation, migration and adhesion.

多肽H2N-Lys-Arg-Ser-Arg-COOH的合成步骤：

1、合成CTC树脂：称取1.9g CTC Resin（如初始取代度约为0.84mmol/g）和1.92mmol Fmoc-Arg(Pbf)-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4.79mmol 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-Arg(Pbf)-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-Arg(Pbf)-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4.79mmol Fmoc-Ser(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9.58mmol DIPEA，4.55mmol 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-Ser(tBu)-Arg(Pbf)-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-Ser(tBu)-Arg(Pbf)-CTC Resin

Fmoc-Arg(Pbf)-Ser(tBu)-Arg(Pbf)-CTC Resin

H2N-Arg(Pbf)-Ser(tBu)-Arg(Pbf)-CTC Resin

Fmoc-Lys(Boc)-Arg(Pbf)-Ser(tBu)-Arg(Pbf)-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-Lys(Boc)-Arg(Pbf)-Ser(tBu)-Arg(Pbf)-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-Lys-Arg-Ser-Arg-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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