Chromium functionalized diglyme plasma polymer coating enhances enzyme-linked immunosorbent assay performance.
Ensuring the optimum orientation, conformation, and density of substrate-bound antibodies is critical for the success of sandwich enzyme-linked immunosorbent assays (ELISAs). In this work, the authors utilize a diethylene glycol dimethyl ether plasma polymer (DGpp) coating, functionalized with chromium within a 96 well plate for the enhanced immobilization of a capture antibody. For an equivalent amount of bound antibody, a tenfold improvement in the ELISA signal intensity is obtained on the DGpp after incubation with chromium, indicative of improved orientation on this surface. Time-of-flight secondary-ion-mass-spectrometry (ToF-SIMS) and principal component analysis were used to probe the molecular species at the surface and showed ion fragments related to lysine, methionine, histidine, and arginine coupled to chromium indicating candidate antibody binding sites. A combined x-ray photoelectron spectroscopy and ToF-SIMS analysis provided a surface molecular characterization that demonstrates antibody binding via the chromium complex. The DGpp+Cr surface treatment holds great promise for improving the efficacy of ELISAs.
Centre for Materials and Surface Science and Department of Chemistry and Physics, La Trobe University, Bundoora VIC 3086, Australia and CSIRO Manufacturing, Clayton, VIC 3168, Australia.
Nicholas G
NG
Welch
Centre for Materials and Surface Science and Department of Chemistry and Physics, La Trobe University, Bundoora VIC 3086, Australia and CSIRO Manufacturing, Clayton, VIC 3168, Australia.
Robert M T
RM
Madiona
CSIRO Manufacturing, Clayton, VIC 3168, Australia.
Christopher D
CD
Easton
CSIRO Manufacturing, Clayton, VIC 3168, Australia.
Judith A
JA
Scoble
Centre for Materials and Surface Science and Department of Chemistry and Physics, La Trobe University, Bundoora, VIC 3086, Australia.
Robert T
RT
Jones
CSIRO Manufacturing, Clayton, VIC 3168, Australia.
Benjamin W
BW
Muir
Centre for Materials and Surface Science and Department of Chemistry and Physics, La Trobe University, Bundoora, VIC 3086, Australia.
Paul J
PJ
Pigram
D000906:Antibodies / Q000378:metabolism*; D001704:Biopolymers / Q000737:chemistry*; D002857:Chromium / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005026:Ethylene Glycols / Q000378:metabolism*; D055767:Immobilized Proteins / Q000378:metabolism*; D008738:Methyl Ethers / Q000378:metabolism*; D018629:Spectrometry, Mass, Secondary Ion
D000906:Antibodies; D001704:Biopolymers; D005026:Ethylene Glycols; D055767:Immobilized Proteins; D008738:Methyl Ethers; D002857:Chromium; C007391:diglyme