Boozer Christina, Ladd Jon, Chen Shengfu, Jiang Shaoyi
Department of Chemical Engineering, University of Washington, Seattle, Washington 98195, USA.
Anal Chem. 2006 Mar 1;78(5):1515-9. doi: 10.1021/ac051923l.
A versatile multichannel biosensor surface is prepared by site-directed immobilization of single-stranded DNA-protein conjugates onto a patterned self-assembled monolayer composed of ssDNA thiols and oligo(ethylene glycol)-terminated thiols. The conjugates each consist of an antibody chemically linked to a unique ssDNA target with a sequence complementary to the surface-bound ssDNA probes and are immobilized on the surface via sequence-specific hybridization. The exceptional specificity of DNA hybridization combined with the diversity of available sequences makes this platform perfect for multichannel sensors. Once the surface is patterned with the appropriate probe sequences, sequence-specific hybridization sorts out the target conjugates and directs them to the appropriate spots on the surface. Previously (Boozer, C. L.; Ladd, J.; Chen, S.; Yu, Q.; Homola, J.; Jiang, S. Anal. Chem. 2004, 76, 6967-6972), we performed proof-of-concept experiments demonstrating the feasibililty of using DNA-directed protein immobilization to produce a single channel biosensor. In this work, we extend this technique and employ DNA-directed protein immobilization to functionalize a multichannel biosensor, which was used for the simultaneous detection of a set of three fertility hormones: human chorionic gonadotropin, human luteinizing hormone, and follicle stimulating hormone by surface plasmon resonance sensor.
通过将单链DNA - 蛋白质共轭物定点固定在由单链DNA硫醇和聚乙二醇末端硫醇组成的图案化自组装单分子层上，制备了一种多功能多通道生物传感器表面。每个共轭物由化学连接到独特单链DNA靶标的抗体组成，该靶标序列与表面结合的单链DNA探针互补，并通过序列特异性杂交固定在表面上。DNA杂交的卓越特异性与可用序列的多样性相结合，使该平台成为多通道传感器的理想选择。一旦表面用适当的探针序列进行图案化，序列特异性杂交就会筛选出目标共轭物，并将它们引导到表面上的适当位置。之前（Boozer, C. L.; Ladd, J.; Chen, S.; Yu, Q.; Homola, J.; Jiang, S. Anal. Chem. 2004, 76, 6967 - 6972），我们进行了概念验证实验，证明了使用DNA定向蛋白质固定来生产单通道生物传感器的可行性。在这项工作中，我们扩展了这项技术，并采用DNA定向蛋白质固定来功能化多通道生物传感器，该传感器通过表面等离子体共振传感器用于同时检测一组三种生育激素：人绒毛膜促性腺激素、人促黄体生成素和卵泡刺激素。
