生物结合
点击化学
叠氮化物
化学
生物正交化学
组合化学
磁性纳米粒子
结合
生物分子
纳米颗粒
溶解
共价键
共轭体系
纳米技术
生物化学
有机化学
材料科学
聚合物
数学分析
数学
作者
G. S. S. Saini,Mrugesh Krishna Parasa,Katherine N. Clayton,Julia G. Fraseur,Scott C. Bolton,Kevin Lin,Steven T. Wereley,Tamara L. Kinzer‐Ursem
标识
DOI:10.1007/s00604-023-06068-4
摘要
Immobilization of proteins and enzymes on solid supports has been utilized in a variety of applications, from improved protein stability on supported catalysts in industrial processes to fabrication of biosensors, biochips, and microdevices. A critical requirement for these applications is facile yet stable covalent conjugation between the immobilized and fully active protein and the solid support to produce stable, highly bio-active conjugates. Here, we report functionalization of solid surfaces (gold nanoparticles and magnetic beads) with bio-active proteins using site-specific and biorthogonal labeling and azide-alkyne cycloaddition, a click chemistry. Specifically, we recombinantly express and selectively label calcium-dependent proteins, calmodulin and calcineurin, and cAMP-dependent protein kinase A (PKA) with N-terminal azide-tags for efficient conjugation to nanoparticles and magnetic beads. We successfully immobilized the proteins on to the solid supports directly from the cell lysate with click chemistry, forgoing the step of purification. This approach is optimized to yield low particle aggregation and high levels of protein activity post-conjugation. The entire process enables streamlined workflows for bioconjugation and highly active conjugated proteins.
科研通智能强力驱动
Strongly Powered by AbleSci AI