4.6 Article

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

Journal

JOURNAL OF CHROMATOGRAPHY A
Volume 1641, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.chroma.2021.461968

Keywords

Analytical instrumentation; Biophysics; Co-eluting proteins; Decay-associated chromatogram; Process analytical technology; Time-resolved intrinsic fluorescence

Funding

  1. Bioprocessing Research Industry Club (BBSRC) [BB/E005942/1]
  2. EPSRC [EP/N025105/1]
  3. UKRI Future Leaders Fellowship [MR/T02156X/1]
  4. BBSRC [BB/E005942/1, BB/K011162/1] Funding Source: UKRI
  5. EPSRC [EP/N025105/1, EP/P006485/1] Funding Source: UKRI

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The adoption of process analytical technologies in the biopharmaceutical industry can reduce the cost of therapeutic drugs and facilitate investigation of new bioprocesses. Control of critical process parameters is crucial for ensuring product quality, but developing the necessary sophisticated analytical technologies has proven to be a challenge.
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. (c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license ( http://creativecommons.org/licenses/by/4.0/ )

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