title: Optimisation of protein microarray techniques for analysis of the plasma proteome: minimisation of non-specific binding interactions.
abstract: Components of the plasma proteome, particularly serum albumin, have been shown to compromise the accuracy of protein microarray technologies through non-specific binding interactions. Optimisation of array conditions is imperative to help address these problems. Here we demonstrate how modifications to array printing conditions and processing methodology can influence the reliability of data output. In particular, we demonstrate that whilst some glycerol is necessary to maintain specific binding signals, it also increases non-specific binding of albumin. Concentrations of 20% glycerol in the printing buffers are therefore recommended. The findings presented here provide opportunities for increased accuracy in plasma protein detection for possible future diagnostic applications.
authors.affiliation: Cell Biophysics Group, Institute of Biophysics, Imaging and Optical Science, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, England, UK.
authors.forename: Joanna L
authors.initials: JL
authors.lastname: Richens
authors.affiliation: Cell Biophysics Group, Institute of Biophysics, Imaging and Optical Science, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, England, UK.
authors.forename: Elizabeth A M
authors.initials: EAM
authors.lastname: Lunt
authors.affiliation: Cell Biophysics Group, Institute of Biophysics, Imaging and Optical Science, School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, England, UK. Electronic address: paul.oshea@nottingham.ac.uk.
authors.forename: Paul
authors.initials: P
authors.lastname: O'Shea
mesh_terms: D000818:Animals; D001798:Blood Proteins / Q000378:metabolism*; D005990:Glycerol / Q000737:chemistry; D006801:Humans; D025202:Molecular Diagnostic Techniques; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D020543:Proteome / Q000378:metabolism*; D058996:Quality Improvement; D015203:Reproducibility of Results
chemical_list: D001798:Blood Proteins; D020543:Proteome; D005990:Glycerol
keywords: Albumin; Glycerol; Microarray; Plasma proteome