COVALENT ATTACHMENT OF AN ARG-GLY-ASP SEQUENCE PEPTIDE TO DERIVATIZABLE POLYACRYLAMIDE SURFACES - SUPPORT OF FIBROBLAST ADHESION AND LONG-TERM GROWTH
COVALENT ATTACHMENT OF AN ARG-GLY-ASP SEQUENCE PEPTIDE TO DERIVATIZABLE POLYACRYLAMIDE SURFACES - SUPPORT OF FIBROBLAST ADHESION AND LONG-TERM GROWTH
复制标题
DOI：
10.1016/0003-2697(88)90442-3
复制
发表时间：
1988-07-01
影响因子：
2.9
通讯作者：
SCHNAAR, RL
中科院分区：
文献类型：
--
作者：
BRANDLEY, BK;SCHNAAR, RL
利用 N-琥珀酰亚胺基活性酯将合成九肽（Tyr-Ala-Val-Thr-Gly-Arg-Gly-Asp-Ser）（包括粘附性 Arg-Gly-Asp (RGD) 序列）共价固定在化学结构明确的聚丙烯酰胺凝胶表面上。固定的肽量随添加到凝胶中的浓度线性变化。固定是.apprx。效率为 80%（基于添加的肽），产生高达 1.75 nmol 的肽/cm2 凝胶表面。即使在没有血清的情况下，Balb/c 3T3 小鼠成纤维细胞也能轻松粘附到肽衍生化表面。此外，用 2 nmol 肽/cm2 凝胶衍生的表面支持成纤维细胞的长期生长，其速度和程度与组织培养塑料上相当。用不具有RGD序列的对照九肽衍生的表面不支持细胞附着或生长。用于用粘附性九肽衍生化凝胶表面的固定技术可以进行修改，以允许与蛋白质、糖蛋白、糖苷或其他含胺化合物进行衍生化，以测试它们对长期细胞行为的影响。
A synthetic nonapeptide (Tyr-Ala-Val-Thr-Gly-Arg-Gly-Asp-Ser), which includes the adhesive Arg-Gly-Asp (RGD) sequence, was covalently immobilized on chemically well-defined polyacrylamide gel surfaces utilizing N-succinimidyl active esters. The amount of peptide immobilized varied linearly with the concentration added to the gels. Immobilization was .apprx. 80% efficient (based on peptide added), resulting in up to 1.75 nmol peptide/cm2 gel surface. Balb/c 3T3 mouse fibroblast cells adhered readily to peptide-derivatized surfaces, even in the absence of serum. Furthermore, surfaces derivatized with 2 nmol peptide/cm2 gel supported long-term fibroblast growth at a rate and to an extent comparable to that on tissue culture plastic. Surfaces derivatized with a control nonapeptide having no RGD sequence were non-supportive of cell attachment or growth. The immobilization technology used to derivatize the gel surfaces with adhesive nonapeptide can be modified to allow conderivatization with proteins, glycoproteins, glycosides, or other amine-containing compounds to test their effects on long-term cell behaviors.