Detection of antigens in biologically complex fluids with photografted whole antibodies

Analytical Chemistry
Robert P SebraKristi S Anseth

Abstract

A highly sensitive (pM), efficient (t < 20 min) detection assay was developed by designing surfaces with grafted antibodies. Through this approach, a short half-life antigen, glucagon, was rapidly detected in a biologically complex plasma/blood environment. Tailoring of graft composition eliminates the need for time-consuming blocking steps, significantly reducing antigen-antibody incubation times, while maintaining antibody specificity and activity toward target antigen. Grafted antibodies were bound through solvated, mobile polymer chains, thereby circumventing problems associated with antibody accessibility, analyte diffusion, and steric limitations. The efficiency of this assay is provided through grafting synthesized, acrylated antibodies in the presence of PEG monoacrylate. This procedure eliminates the need for blocking steps, due to a decrease in nonspecific protein interactions. These polymerizable antibodies were tethered with a range of densities while retaining biological activity. Moreover, biological activity of acrylated antibodies was compared to that of unmodified antibodies and remained comparable. The acrylated antibodies were grafted from substrate surfaces using controlled radical photopolymerization, maint...Continue Reading

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Citations

Apr 28, 2010·Analytical and Bioanalytical Chemistry·Alphonsus H C NgAaron R Wheeler
Sep 29, 2011·International Journal of Molecular Sciences·Andreas E Vasdekis, Gregoire P J Laporte
Nov 19, 2010·Bioanalysis·Michael C Peoples, H Thomas Karnes
Sep 5, 2013·Biomicrofluidics·Dohyun Kim, Amy E Herr
Mar 2, 2010·Clinical Orthopaedics and Related Research·McKinley C LawsonKristi S Anseth
Jul 13, 2007·Biotechnology and Bioengineering·Angela T LewandowskiWilliam E Bentley

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