A label‐free methodology for selective protein quantification by means of absorption measurements

Hansen, Sigrid K.; Skibsted, Erik; Staby, Arne; Hubbuch, Jürgen

doi:10.1002/bit.23229

Cited by 35 publications

(31 citation statements)

References 18 publications

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“…In summary, the prediction accuracy of the developed PLS model for quantifying the protein mixtures was acceptable owing to the use of multi‐wavelength UV spectra, and their validity was rigorously analyzed using the various kinds of chemometrics tools. In addition, it is worth to note that the approach employed here was different from the previous studies, where a single type of mixture design has been employed without considering the use of fully independent test set. Table summarized the advantages of the proposed approach in terms of chromatographic operation for protein purification, compared to other methodologies reported in the literature .…”

Section: Resultsmentioning

confidence: 99%

“…In this study, the feasibility of multiwavelength UV absorption spectra was investigated to estimate the protein concentrations in mixture as a potential real‐time quantitative monitoring tool in protein purification, considering that it can be readily implemented in many of the existing chromatography systems with minimal modification and has been well‐established techniques with maximal robustness and reproducibility . In fact, several researchers have previously reported the successful applications of UV spectroscopy in analyzing the structural behavior of the protein or in quantifying the composition of proteins mixtures . However, the majority of them have still used a univariate or a few wavelengths of high importance (i.e., ratio of two absorbance, etc.…”

Section: Introductionmentioning

confidence: 99%

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Quantification of protein mixture in chromatographic separation using multi‐wavelength UV spectra

Kamga

Lee

Liu

et al. 2013

Biotechnology Progress

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In therapeutic protein production, the protein purification with chromatographic processes is of high importance in separating the qualified proteins from the impurities for consistent product quality. Therefore, to aid real-time monitoring of the protein purification processes, various kinds of methodologies have been proposed until now. However, the majority of them still rely on the use of a single ultraviolet (UV) absorbance or the utilization of expensive and time-consuming instruments, thus requiring a simple, fast, and cost-effective methodology for protein quantification. In this study, the feasibility of using multiwavelength UV spectroscopy was investigated as an alternative tool for the real-time monitoring of the protein mixtures in protein purification. To this end, three different proteins were selected as a model system for the protein mixture, and the multivariate UV spectra were analyzed to construct the reliable quantification models for different proteins of interest. By using various chemometrics tools, such as partial least squares (PLS), the validity of estimating the protein concentration from the UV spectra of the mixture samples was rigorously analyzed with their prediction performance, and the results indicated that the multiwavelength UV spectra had sufficient sensitivity and accuracy to estimate the protein concentrations in mixture, demonstrating its usefulness for the rapid quantification of the protein mixtures in protein purification.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Quantification of protein mixture in chromatographic separation using multi‐wavelength UV spectra

Kamga

Lee

Liu

et al. 2013

Biotechnology Progress

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“… used near‐infrared spectral data for investigating complex feedstocks from an antibiotic production process. Deconvolution of protein spectra in the mid‐UV region was shown to be highly useful in determining protein concentrations from multi‐component mixtures . Brestrich et al.…”

Section: Modeling Approachesmentioning

confidence: 99%

Downstream process development strategies for effective bioprocesses: Trends, progress, and combinatorial approaches

Baumann

Hubbuch

2016

Engineering in Life Sciences

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The biopharmaceutical industry is at a turning point moving toward a more customized and patient-oriented medicine (precision medicine). Straightforward routines such as the antibody platform process are extended to production processes for a new portfolio of molecules. As a consequence, individual and tailored productions require generic approaches for a fast and dedicated purification process development. In this article, different effective strategies in biopharmaceutical purification process development are reviewed that can analogously be used for the new generation of antibodies. Conventional approaches based on heuristics and high-throughput process development are discussed and compared to modern technologies such as multivariate calibration and mechanistic modeling tools. Such approaches constitute a good foundation for fast and effective process development for new products and processes, but their full potential becomes obvious in a correlated combination. Thus, different combinatorial approaches are presented, which might become future directions in the biopharmaceutical industry.

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“…Moreover, the feed, with concentration c load,α and α ∈ {A, B,C}, is injected as a rectangular pulse with duration ∆t load via a multiport valve. The eluting concentration trajectory is analyzed with a UV detector at the column outlet [54], and a fractionating valve separates the mixture into its components [36]. The fractionating interval manipulating variables [τ 0 , ∆τ] govern the maximum recovery yield while still fulfilling the constraint imposed on purity of the target component fraction [16].…”

Section: Process Descriptionmentioning

confidence: 99%

Methods and Tools for Robust Optimal Control of Batch Chromatographic Separation Processes

et al. 2015

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This contribution concerns the development of generic methods and tools for robust optimal control of high-pressure liquid chromatographic separation processes. The proposed methodology exploits a deterministic robust formulation, that employs a linearization of the uncertainty set, based on Lyapunov differential equations to generate optimal elution trajectories in the presence of uncertainty. Computational tractability is obtained by casting the robust counterpart problem in the framework of bilevel optimal control where the upper level concerns forward simulation of the Lyapunov differential equation, and the nominal open-loop optimal control problem augmented with the robustified target component purity inequality constraint margin is considered in the lower level. The lower-level open-loop optimal control problem, constrained by spatially discretized partial differential equations, is transcribed into a finite dimensional nonlinear program using direct collocation, which is then solved by a primal-dual interior point method. The advantages of the robustification strategy are highlighted through the solution of a challenging ternary complex mixture separation problem for a hydrophobic interaction chromatography system. The study shows that penalizing the changes in the zero-order hold control gives optimal solutions with low sensitivity to uncertainty. A key result is that the robustified general elution trajectories outperformed the conventional linear trajectories both in terms of recovery yield and robustness.

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A label‐free methodology for selective protein quantification by means of absorption measurements

Cited by 35 publications

References 18 publications

Quantification of protein mixture in chromatographic separation using multi‐wavelength UV spectra

Quantification of protein mixture in chromatographic separation using multi‐wavelength UV spectra

Downstream process development strategies for effective bioprocesses: Trends, progress, and combinatorial approaches

Methods and Tools for Robust Optimal Control of Batch Chromatographic Separation Processes

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