Intrinsic fluorescence-basedat situsoft sensor for monitoring monoclonal antibody aggregation
- 22 July 2015
- journal article
- research article
- Published by Wiley in Biotechnology Progress
- Vol. 31 (5), 1423-1432
- https://doi.org/10.1002/btpr.2140
Abstract
Intrinsic fluorescence spectroscopy, in conjunction with partial least squares regression (PLSR), was investigated as a potential technique for online quality control and quantitative monitoring of Immunoglobulin G (IgG) aggregation that occurs following exposure to conditions that emulate those that can occur during protein downstream processing. Initially, the impact of three stress factors (temperature, pH, and protein concentration) on the degree of aggregation determined using size exclusion chromatography data, was investigated by performing a central composite designexperiment and applying a fitting response surface model. This investigation identified the influence of the factors as well as the operating regions with minimum propensity to induce protein aggregation. Spectral changes pertinent to the stressed samples were also investigated and found to corroborate the high sensitivity of the intrinsic fluorescence to conformational changes of the proteins under study. Ultimately, partial least squares regression was implemented to formulate two fluorescence-based soft sensors for quality control--product classification--and quantitative monitoring--concentration of monomer. The resulting regression models exhibited accurate prediction ability and good potential for in situ monitoring of monoclonal antibody downstream purification processes.Keywords
Funding Information
- Natural Science and Engineering Research Council (NSERC)
- Monoclonal Antibody Network (MabNet)
This publication has 21 references indexed in Scilit:
- Comparative Effects of pH and Ionic Strength on Protein–Protein Interactions, Unfolding, and Aggregation for IgG1 AntibodiesJournal of Pharmaceutical Sciences, 2010
- Detection of IgG Aggregation by a High Throughput Method Based on Extrinsic FluorescenceJournal of Pharmaceutical Sciences, 2010
- Advances in on-line monitoring and control of mammalian cell cultures: Supporting the PAT initiativeBiotechnology Advances, 2009
- Protein aggregation: Pathways, induction factors and analysisJournal of Pharmaceutical Sciences, 2009
- A simple and reproducible approach to characterize protein stability using rheologyInternational Journal of Pharmaceutics, 2009
- Fluorescence-based soft-sensor for monitoring β-lactoglobulin and α-lactalbumin solubility during thermal aggregationBiotechnology & Bioengineering, 2007
- Fluorescence spectroscopy as a tool for monitoring solubility and aggregation behavior of β‐lactoglobulin after heat treatmentBiotechnology & Bioengineering, 2006
- Second derivative tryptophan fluorescence spectroscopy as a tool to characterize partially unfolded intermediates of proteinsInternational Journal of Pharmaceutics, 2005
- Protein aggregation and its inhibition in biopharmaceuticsInternational Journal of Pharmaceutics, 2005
- Partial least-squares regression: a tutorialAnalytica Chimica Acta, 1986