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首页> 外文期刊>Biotechnology Progress >Cytophobic Surface Modification of Microfluidic Arrays for In Situ Parallel Peptide Synthesis and Cell Adhesion Assays
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Cytophobic Surface Modification of Microfluidic Arrays for In Situ Parallel Peptide Synthesis and Cell Adhesion Assays

机译：微流控阵列的疏水表面修饰，用于原位平行肽合成和细胞粘附测定

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A combination of PEG-based surface passivation techniques and spatially addressable SPPS (solid-phase peptide synthesis) was used to demonstrate a highly specific cell-peptide adhesion assay on a microfluidic platform.The surface of a silicon-glass microchip was modified to form a mixed self-assembled monolayer that presented PEG moieties interspersed with reactive amino terminals.The PEG provided biomolecular inertness and the reactive amino groups were used for consequent peptide synthesis.The cytophobicity of the surface was characterized by on-chip fluorescent binding assays and was found to be resistant to nonspecific attachment of cells and proteins.An integrated system for parallel peptide synthesis on this reactive amino surface was developed using photogenerated acid chemistry and digital microlithography.A constant synthesis efficiency of >98% was observed for up to 7mer peptides.To demonstrate specific cell adhesion on these synthetic peptide arrays,variations of a 7mer cell binding peptide that binds to murine B lymphoma cells were synthesized.Sequence-specific binding was observed on incubation with fluorescently labeled,intact murine B lymphoma cells,and key residues for binding were identified by deletional analysis.
机译：结合使用基于PEG的表面钝化技术和空间可寻址SPPS（固相肽合成）技术，在微流控平台上展示了​​高度特异性的细胞-肽粘附测定法。混合的自组装单层结构，其PEG部分散布在反应性氨基末端上.PEG提供了生物分子惰性，反应性氨基用于随后的肽合成。通过片上荧光结合试验表征了表面的疏油性，发现利用光生酸化学和数字微光刻技术开发了在此反应性氨基表面上平行肽合成的集成系统，在多达7个肽的情况下，合成效率恒定在98％以上。这些合成肽阵列上的特异性细胞粘附，7mer细胞的变异合成了与鼠B淋巴瘤细胞结合的结合肽。与荧光标记的完整鼠B淋巴瘤细胞一起孵育时观察到序列特异性结合，并通过缺失分析鉴定了结合的关键残基。

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