Abstract

Photo-Electric Microbe Sensor is a patented biotechnology that detects microbes in aqueous solution by measuring the change in photo-voltage in response to UV light stimulation of a platinum (Pt) disk surface on an electrode before and after immunoprecipitation of microbes. This study aimed to increase the sensitivity of microbe detection by pre-adsorbing recombinant Streptococcal Protein G (PG), to the Pt sensor surface. Streptococcal PG binds the Fc region of mammalian IgG molecules and we investigated the association of PG to Pt and the resulting ability to tether antibodies to the Pt-PG surface. An ELISA protocol was optimized to detect the presence of mouse monoclonal antibodies tethered to Pt immunoaffinity disks, and to determine the recommended blocking solution and reagent concentrations. Our results demonstrate that PG binds to bare Pt, increases IgG affinity to the Pt surface following Superblock Buffer application, and together offers design-options for Pt-based sensor technologies.

Highlights

  • Introduction(2015) Streptococcal Protein G Enhances Antibody Binding to Platinum Sensor Surfaces

  • Superblock Blocking Buffer in phosphate buffered saline (PBS), recombinant Streptococcal Protein G (PG), Mouse IgG anti-lipoteichoic acid (LTA), and Rabbit anti-Mouse IgG-horse radish-peroxidase (HRP) conjugate were purchased from Pierce Antibodies, Thermo Scientific

  • In an effort to enhance antibody binding to metal surfaces of Photo-Electric Microbe Sensor (PEMS) electrodes, PG was bound to platinum immunoaffinity disks (Pt disk) by exposure to varying concentrations of recombinant PG

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Summary

Introduction

(2015) Streptococcal Protein G Enhances Antibody Binding to Platinum Sensor Surfaces. (a) PEMs electrode: Immunoaffinity Pt disk mounted on dual circuit board electrode prototype. Three arrows indicate the locations of leads to electrometer, the Pt disk providing an available surface (3 mm diameter) for PG and antibody treatment and the UV-Vis cuvette containing solution of target antigens or molecules. The enlarged graphic diagram of electrodes indicates dual Pt surfaces and surrounding charges when submerged in 1 mM NaCl test solution. (b) The strobe generates light in the UV spectra which is focused through a series of lenses onto a Pt disk. The subsequent movement of charge produces a photovoltage that is amplified by the electrometer and displayed on an oscilloscope

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