Optimisation of protein microarray techniques for analysis of the plasma proteome: Minimisation of non-specific binding interactions |
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| Affiliation: | 1. State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, China;2. Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;3. Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, College of Life Science, Jilin University, Changchun 130012, China;1. Shanghai Key Lab of Chemistry Biology, School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China;2. Shanghai Collaborative Innovation Center for Biomanufacturing Technology, Shanghai 200237, China;1. Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, 201800, China;2. Shanghai Synchrotron Radiation Facility, Shanghai Advanced Research Institute, Chinese Academy of Sciences, Shanghai, 201204, China;3. University of Chinese Academy of Sciences, Beijing, 100049, China;1. H. H. Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol, BS8 1TL, UK;2. DSM Nutritional Products AG, Research Center Formulation & Application, P.O. Box 2676, 4002 Basel, Switzerland;3. Bristol Centre for Functional Nanomaterials, H. H. Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL, UK;4. Food Concept & Physical Design “The Mill”, Mühleweg 10, CH-4112 Flüh, Switzerland |
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| 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. |
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