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<p>In this study, an amine-reactive poly(pentafluorophenyl acrylate) (<b>PPFPA</b>) platform was developed for advanced surface engineering of next-generation sequencing (NGS) chips. Through post-polymerization modification, <b>PPFPA</b> was functionalized with dual moieties: azide groups for covalent immobilization of DBCO-modified DNA primers <i>via</i> click chemistry and tunable hydrophilic side chains to optimize biocompatibility and surface properties. Systematic screening revealed that hydrophobic azide carriers combined with neutral hydroxyl groups maximized the DNA immobilization efficacy, approaching the performance of commercial polyacrylamide-based polymers. The negatively charged carboxyl groups severely impede DNA primer attachment. Higher molecular weight derivatives further enhance the efficacy of DNA immobilization. In NGS validation, optimized surface modification polymers achieved robust surface density of clustered DNA and high sequencing accuracy, surpassing quality benchmarks and comparable to those of conventional analogs. This platform demonstrates significant potential for tailoring high-sensitivity surfaces for genomic applications, advancing clinical diagnostics, and personalized medicine.</p>

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Amine-reactive Polymer Platform for Engineering Surface Modification of Next-generation Sequencing Chips

  • 期刊论文

  • 发表日期：

    2025年9月2日

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  • Wei Tian,
  • Xin-Yuan Wang,
  • Die-Wen Feng,
  • Xiang-Qian Li,
  • Yue-Kang Jin,
  • Hui Li,
  • Hao Liu

摘要

In this study, an amine-reactive poly(pentafluorophenyl acrylate) (PPFPA) platform was developed for advanced surface engineering of next-generation sequencing (NGS) chips. Through post-polymerization modification, PPFPA was functionalized with dual moieties: azide groups for covalent immobilization of DBCO-modified DNA primers via click chemistry and tunable hydrophilic side chains to optimize biocompatibility and surface properties. Systematic screening revealed that hydrophobic azide carriers combined with neutral hydroxyl groups maximized the DNA immobilization efficacy, approaching the performance of commercial polyacrylamide-based polymers. The negatively charged carboxyl groups severely impede DNA primer attachment. Higher molecular weight derivatives further enhance the efficacy of DNA immobilization. In NGS validation, optimized surface modification polymers achieved robust surface density of clustered DNA and high sequencing accuracy, surpassing quality benchmarks and comparable to those of conventional analogs. This platform demonstrates significant potential for tailoring high-sensitivity surfaces for genomic applications, advancing clinical diagnostics, and personalized medicine.