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    Recombinant Spider Silk as Mediator for One-Step, Chemical-Free Surface Biofunctionalization

    Year: 2018

    Journal: Adv. Funct. Mater., Volume 28, MAY 24

    Authors: Horak, Josef; Jansson, Ronnie; Dev, Apurba; Nileback, Linnea; Behnam, Kiarash; Linnros, Jan; Hedhammar, My; Karlstrom, Amelie Eriksson

    Keywords: biomarker; biosensing; C1q; surface biofunctionalization; Z-silk

    A unique strategy for effective, versatile, and facile surface biofunctionalization employing a recombinant spider silk protein genetically functionalized with the antibody-binding Z domain (Z-4RepCT) is reported. It is demonstrated that Z-silk can be applied to a variety of materials and platform designs as a truly one-step and chemical-free surface modification that site specifically captures antibodies while simultaneously reducing nonspecific adsorption. As a model surface, SiO2 is used to optimize and characterize Z-silk performance compared to the Z domain immobilized by a standard silanization method. First, Z-silk adsorption is investigated and verified its biofunctionality in a long-term stability experiment. To assess the binding capacity and protein-protein interaction stability of Z-silk, the coating is used to capture human antibodies in various assay formats. An eightfold higher binding capacity and 40-fold lower detection limit are obtained in the immunofluorescence assay, and the complex stability of captured antibodies is shown to be improved by a factor of 20. Applicability of Z-silk to functionalize microfluidic devices is demonstrated by antibody detection in an electrokinetic microcapillary biosensor. To test Z-silk for biomarker applications, real-time detection and quantification of human immunoglobulin G are performed in a plasma sample and C1q capture from human serum using an anti-C1q antibody.
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