Automated measurement of apparent protein solubility to rapidly assess complex parameter interactions

Berg, Annette; Schuetz, Maren; Dismer, Florian; Hubbuch, Jürgen

doi:10.1016/j.fbp.2013.10.002

Cited by 11 publications

(10 citation statements)

References 33 publications

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“…Despite the availability of various stabilizing additives, identification of the most effective additive from a large experimental space for each target protein is laborious and timeconsuming using analytical methods with low-throughput capacity [41,42]. Therefore, the use of high-throughput screening (HTS) techniques may provide opportunities for simple and rapid screening for the most effective additives.…”

Section: Introductionmentioning

confidence: 99%

“…HTS techniques have become a valuable tool for speeding up process development at various stages of processing biopharmaceuticals [43,44]. They have been used effectively for rapid identification and selection of conditions for precipitation [45] and separation [46] of monoclonal antibodies from host cell proteins; for determining protein solubility [41,43] and rapid assessment of the dependence of colloidal stability on complex parameter interactions [41]; and for development of production processes [47] and quantification [48] of polysaccharide vaccine candidates. Mohr et al [40] demonstrated a miniaturized HTS methodology for VLP formulation by integrating dynamic light scattering (DLS) and asymmetrical flow field-flow fractionation.…”

Section: Introductionmentioning

confidence: 99%

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A rapid and simple screening method to identify conditions for enhanced stability of modular vaccine candidates

Tekewe

Connors

Sainsbury

et al. 2015

Biochemical Engineering Journal

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A rapid and simple screening method to identify conditions for enhanced stability of modular vaccine candidates

Tekewe

Connors

Sainsbury

et al. 2015

Biochemical Engineering Journal

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“…It indicates that S Ly decreases along with the increase of c NaCl and reaches a low limit. Compared with Ly solubility data measured by Berg et al 43 and Forsythe et al 50 under the same conditions, Figure 4 indicates that the solubility curve measured by the UDDCS method is located in the middle of the other two curves.…”

Section: Resultsmentioning

confidence: 71%

Development of a Method for Fast Assessment of Protein Solubility Based on Ultrasonic Dispersion and Differential Centrifugation Technology

Wei

Wang

et al. 2022

ACS Omega

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Protein solubility is very important for protein crystallization, bioprocess development, and protein application. In this study, a method based on the stability of a protein dispersion system is proposed for fast assessment of protein solubility, which mainly involves ultrasonic dispersion, differential centrifugation, and spectral measurement (UDDCS) and curvature estimation. The appropriate ultrasonic time and centrifugal time were experimentally determined at first. The results show that the relationship between the standard deviation and the protein concentrations originally added accords with the modified exponential equation, and the corresponding concentration of the maximum curvature point is defined as the solubility of the protein. Lysozyme solubility data in NaCl aqueous solutions and zein solubility data in ethanol aqueous solutions are selected to verify the UDDCS method by comparing the data obtained by the UDDCS method and the results from references, and the results indicate that the UDDCS method is reliable, universal, and time-saving. Finally, measurements of zein solubility in NaOH solution and casein solubility in urea aqueous solution were conducted as test cases by the UDDCS method.

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“…RNAse A was found to be destabilized by moderate sodium chloride concentrations of 250 and 500 mM . Lysozyme also showed a strong decrease in solubility at salt concentrations below 0.5 M .…”

Section: Introductionmentioning

confidence: 98%

“…Reducing agents, such as DTT in low concentrations, were applied as well . Lysozyme is shown to be stabilized by low molecular weight PEGs, detergents, such as Tween 20, and different sugar molecules such as sorbitol and sucrose .…”

Section: Introductionmentioning

confidence: 99%

Systematic purification of salt‐intolerant proteins by ion‐exchange chromatography: The example of human α‐galactosidase A

Baumann

Osberghaus

Hubbuch

2015

Engineering in Life Sciences

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Chromatography is an essential tool for purifying biopharmaceutical products. Many processes are still developed based on traditional routines and empirical procedures. Product losses are mostly due to insufficient optimization of purification setups and product sensitivity to process conditions. In order to eliminate these shortcomings, a systematic strategy for the setup of ion‐exchange chromatography is presented, which considers both product stability as well as operational conditions. The stages—a hybrid approach combining high‐throughput screening and analytical small‐scale chromatography—are as follows: (1) pH stability (short‐term); (2) pH stability (long‐term), followed by a screening of additives to enhance protein stability, if required; (3) analytical pH gradient chromatography for evaluation of the operational pH window; and (4) salt stability (long‐term) in the operational pH window determined. The efficiency and straightforwardness of the strategy were shown in a case study on capturing the human α‐galactosidase A enzyme. Following the above procedure, the enzyme was found to be salt‐unstable; a purification factor of 13.2, a concentration factor of 4, and an overall yield of 84.3% were achieved. The applied strategy allowed for a quick establishment of a dedicated capture step at low salt concentrations under stable conditions by well‐chosen prior screening experiments.

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Automated measurement of apparent protein solubility to rapidly assess complex parameter interactions

Cited by 11 publications

References 33 publications

A rapid and simple screening method to identify conditions for enhanced stability of modular vaccine candidates

A rapid and simple screening method to identify conditions for enhanced stability of modular vaccine candidates

Development of a Method for Fast Assessment of Protein Solubility Based on Ultrasonic Dispersion and Differential Centrifugation Technology

Systematic purification of salt‐intolerant proteins by ion‐exchange chromatography: The example of human α‐galactosidase A

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