Immobilization of Active Antibodies at Polymer Melt Surfaces during Injection Molding

Abstract

We demonstrate the transfer and immobilization of active antibodies from a low surface- energy mold surface to thermoplastic replica surfaces using injection molding, and we investigate the process at molecular scale. The transfer process is highly efficient, as verified by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) of the mold and replica surfaces. AFM analysis reveals partial nanometer-scale embedding of the protein into the polymer matrix as a possible mechanism of permanent immobilization. Replicas with rabbit anti-mouse IgG immobilized as capture antibody at the hot polymer melt surface during injection molding show similar affinity for their antigen (mouse IgG) in sandwich enzyme-linked immunosorbent assay (ELISA) as capture antibodies deposited by passive adsorption onto a bare thermoplastic replica. The transferred antibodies retain their functionality after incubation in serum-containing cell medium for >1 week. A mold coating time of 10 min prior to injection molding is sufficient for producing highly sensitive ELISA assays, thus enabling the short processing cycle times required for mass production of single-use biodevices relying on active immobilized antibodies.