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Proof-of-concept analytical instrument for label-free optical de-convolution of protein species in a mixture

Hales, JE; Aoudjane, S; Aeppli, G; Dalby, PA; (2021) Proof-of-concept analytical instrument for label-free optical de-convolution of protein species in a mixture. Journal of Chromatography A , 1641 , Article 461968. 10.1016/j.chroma.2021.461968. Green open access

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Abstract

The adoption of process analytical technologies by the biopharmaceutical industry can reduce the cost of therapeutic drugs and facilitate investigation of new bioprocesses. Control of critical process parameters to retain critical product quality attributes within strict bounds is important for ensuring a consistently high product quality, but developing the sophisticated analytical technologies required has proven to be a major challenge. Here, we demonstrate a new optical technique for continuous monitoring of protein species as they are eluted from a chromatographic column, even when they fully co-elute with other protein species, without making any assumption about or peak-fitting to the elution profile. To achieve this, we designed and constructed a time-resolved intrinsic fluorescence lifetime chromatograph, and established an analytical framework for deconvolving and quantifying distinct but co-eluting protein species in real time. This proof-of-concept technology has potentially useful applications as a process analytical technology and more generally as an analytical technique for label-free quantification of proteins in mixtures.

Type: Article
Title: Proof-of-concept analytical instrument for label-free optical de-convolution of protein species in a mixture
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.chroma.2021.461968
Publisher version: https://doi.org/10.1016/j.chroma.2021.461968
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Analytical instrumentation, Biophysics, Co-eluting proteins, Decay-associated chromatogram, Process analytical technology, Time-resolved intrinsic fluorescence.
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10121405
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