Generation of Equally Sized Particle Plaques Using Solid-Liquid Suspensions

Herrmann, Tim; Schröder, Magnus; Hubbuch, Jürgen

doi:10.1021/bp050296i

Cited by 30 publications

(21 citation statements)

References 7 publications

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“…Finally, pipetting of slurries expresses a high potential for experimental errors. In contrast, the use of resin plaques of defined volume prepared prior to an experimental study has shown to express a superior performance and eliminates the above mentioned drawbacks [28]. Plaques are generated using a grid in 96-well plate format with holes of a defined volume.…”

Section: Plate Handlingmentioning

confidence: 99%

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High Throughput Screening of Chromatographic Phases for Rapid Process Development

et al. 2005

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The necessity of being ªfirst on the marketº paired with the need to cut production costs highlights the need for strategies and techniques allowing a fast process development while at the same time covering a high number of potential process parameters in order to reach an optimal process sequence. This can only be met by the use of high throughput screening (HTS) techniques. In this paper HTS applications presently applied in bioprocess development are reviewed, their use in chromatographic process parameter evaluation is discussed and concepts for the rational design of HTS experiments are presented. A rational approach for chromatographic parameter screening comprehends an initial screening procedure for ligand selection followed by the incorporation of specific adsorbent features using HTS pipetting systems finalized by a fine tuning step using packed mini-columns. The analytical part of these screening techniques clearly sets the bottleneck of these procedures with a trade off between analytical information and analysis time. Finally, intelligent HTS strategies should in addition include efficient data processing paired with modern tools for design of experiments or model based process design.

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Section: Plate Handlingmentioning

confidence: 99%

“…Plaques are generated using a grid in 96-well plate format with holes of a defined volume. The holes are then filled with the respective resins by using vacuum [28] or centrifugal force [29].…”

Section: Plate Handlingmentioning

confidence: 99%

High Throughput Screening of Chromatographic Phases for Rapid Process Development

et al. 2005

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“…A vacuum plaque device was used to generate equally sized adsorbent plaques with a volume of 8.8 L each (20). The plaques were equilibrated within the plaque device with 100 mM Tris-HCl buffer, pH 8.0.…”

Section: Methodsmentioning

confidence: 99%

Lectin-Based Affinity Tag for One-Step Protein Purification

et al. 2006

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The production of pure protein is indispensable for many applications in life sciences, however protein purification protocols are difficult to establish, and the experimental procedures are usually tedious and time-consuming. Therefore, a number of tags were developed to which proteins of interest can be fused and subsequently purified by affinity chromatography. We report here on a novel lectin-based affinity tag using the D-mannose-specific lectin LecB from Pseudomonas aeruginosa. A fusion protein was constructed consisting of yellow fluorescent protein and LecB separated by an enterokinase cleavage site. This protein was overexpressed in Escherichia coli Tuner (DE3), and the cell extract was loaded onto a column containing a mannose agarose matrix. Electrophoretically pure fusion protein at a yield of 24 mg/L culture was eluted with a D-mannose containing buffer The determination of equilibrium adsorption isotherms revealed an association constant of the lectin to the mannose agarose matrix of Ka = 3.26 x 10(5)/M. Enterokinase treatment of the purified fusion protein resulted in the complete removal of the LecB-tag. In conclusion, our results indicate that the lectin LecB of P. aeruginosa can be used as a tag for the high-yield one-step purification of recombinant proteins.

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“…As the 36 plaques at the border of the ResiQuot device show slightly higher variances in shape and weight (see [18]), they were completely left out in experimentation, thus only the 60 inner plaques in the deep-well plate (DWP) were used for the binding studies. S1a in the Supporting Information illustrates the size and appearance of typical adsorbent plaques.…”

Section: Apparatus and Softwarementioning

confidence: 99%

Detection, Quantification, and Propagation of Uncertainty in High‐Throughput Experimentation by Monte Carlo Methods

et al. 2012

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Since the efficiency and speed of computing has increased significantly in the last decades, in silico‐approaches, e.g., quasi‐experimental analyses based on mechanistic simulations combined with Monte Carlo (MC) methods, are on the rise for uncertainty analyses and estimation of uncertainty propagation. The power and convenience of these approaches for high‐throughput processes will be demonstrated with a case study including miniaturized screenings on robotic platforms: a binding study for lysozyme on the adsorbent SP Sepharose FF in 96‐well format. All relevant uncertainties during the experimental preparations and automated high‐throughput experimentation were identified, quantified, and then embedded in a simulation algorithm for the calculation of uncertainty propagation based on MC sampling. A proof‐of‐concept for this approach is then followed by the simulation‐based analysis of various case scenarios. The MC‐based approach can easily be transferred to uncertainty analyses in other high‐throughput processes.

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Generation of Equally Sized Particle Plaques Using Solid-Liquid Suspensions

Cited by 30 publications

References 7 publications

High Throughput Screening of Chromatographic Phases for Rapid Process Development

High Throughput Screening of Chromatographic Phases for Rapid Process Development

Lectin-Based Affinity Tag for One-Step Protein Purification

Detection, Quantification, and Propagation of Uncertainty in High‐Throughput Experimentation by Monte Carlo Methods

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