title: High-Throughput Production of Chromium(III) Complexes for Antibody Immobilization.
abstract: A robot-assisted high-throughput methodology was employed to produce chromium(III) complexes suitable for the surface modification of the commercially available PerkinElmer Optiplate96 well plate for use in enzyme-linked immunosorbent assays (ELISAs). The complexes were immobilized to the native functionality of the well plate and first screened using a horseradish-peroxidase-tagged (HRP) mouse antibody to quantify binding. The top "hits" were further assessed for their ability to present the antibody in a functional state using an ELISA. "Hits" from the second screen yielded four complexes capable of improving the signal intensity of the ELISA by greater than 500%. The metal/base ratio of these complexes was also investigated, and we isolated the most stable and reproducible candidate, [Cr(OH)6]3-, which was formed from chromium(III) perchlorate and pH adjusted with ethylenediamine. This chromium solution was employed in a clinically relevant setting for the detection of bovine TNFα producing up to a 200% increase in signal intensity.
authors.affiliation: Centre for Materials and Surface Science and Department of Chemistry and Physics, La Trobe University , Bundoora, Victoria 3086, Australia.|CSIRO, Manufacturing, Clayton, Victoria 3168, Australia.
authors.forename: Nicholas G
authors.initials: NG
authors.lastname: Welch
authors.affiliation: CSIRO, Manufacturing, Clayton, Victoria 3168, Australia.
authors.forename: Judith A
authors.initials: JA
authors.lastname: Scoble
authors.affiliation: CSIRO, Manufacturing, Clayton, Victoria 3168, Australia.
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Easton
authors.affiliation: CSIRO, Manufacturing, Clayton, Victoria 3168, Australia.
authors.forename: Charlotte C
authors.initials: CC
authors.lastname: Williams
authors.affiliation: Kansas State University , Department of Animal Sciences and Industry, Manhattan, Kansas 66506, United States.
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Bradford
authors.affiliation: Kansas State University , Department of Animal Sciences and Industry, Manhattan, Kansas 66506, United States.
authors.forename: Laman K
authors.initials: LK
authors.lastname: Mamedova
authors.affiliation: Centre for Materials and Surface Science and Department of Chemistry and Physics, La Trobe University , Bundoora, Victoria 3086, Australia.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Pigram
authors.affiliation: CSIRO, Manufacturing, Clayton, Victoria 3168, Australia.
authors.forename: Benjamin W
authors.initials: BW
authors.lastname: Muir