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Towards the development of protein-protein interaction standards to improve the comparability of protein interaction measurements.

Moore, J D; Noble, J E; Nugent, P G; Cerasoli, E; Lamarre, B M; Bailey, M J A (2007) Towards the development of protein-protein interaction standards to improve the comparability of protein interaction measurements. NPL Report. AS 8

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Abstract

In order to improve the inter-platform comparability of protein-protein interaction measurements, there is a need for appropriate protein-protein interaction model systems to be developed and assessed. This report summarises work carried out, under the Measurements for Biotechnology (MfB) Programme 2004-2007, towards the development and characterisation of such model systems. By reviewing the needs for the measurement of protein-protein interactions, two model systems were selected, the E. coli multi-subunit enzyme Tryptophan Synthase, and the E. coli N Utilisation substance A / Phage Lambda N antitermination protein interaction pairs, were selected as suitable model systems. These were produced in-house, and assessed using two industrially important platforms: isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR). Published protein expression and purification protocols were modified to improve their robustness and the binding activity of each interaction pair was successfully characterised using the two orthogonal measurement platforms. The values for binding affinity, generated using the two techniques, agreed well. It was found, however, that in the case of SPR, this agreement was dependent on the surface immobilisation strategy used. Both interaction pairs were assessed as potential protein-protein interaction standards and it was concluded that only the TrpA/TrpB pair appeared suitable.

Item Type: Report/Guide (NPL Report)
NPL Report No.: AS 8
Keywords: surface plasma resonance, isothermal titration calorimetry, protein-protein interaction
Subjects: Biotechnology
Biotechnology > Biopharmaceutical Manufacturing and Characterisation
Last Modified: 02 Feb 2018 13:15
URI: http://eprintspublications.npl.co.uk/id/eprint/3923

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