Engineering characterisation of a single well from 24-well and 96-well microtitre plates

Zhang, Hu; Lamping, Sally R.; Pickering, Samuel C.R.; Lye, Gary J.; Shamlou, P. Ayazi

doi:10.1016/j.bej.2007.12.005

Cited by 66 publications

(67 citation statements)

References 29 publications

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“…While experimental assessment of P/V in shaken microwells is currently not feasible, CFD simulations of microwells can be used to predict P/V, and various other relevant parameters such as shear rate. We have previously applied CFD to both shaken flasks (Zhang et al, 2005) and other microwell geometries (Zhang et al, 2008) and the approach has recently been adopted by other groups (Zhang et al, 2009). CFD simulations of the shear rate in shaken 24-SRW microtiter plates as used here are shown in Figure 4.…”

Section: Characterization Of Mean Energy Dissipation Ratesmentioning

confidence: 98%

“…The volume of fluid (VOF) method was used to solve the transient forms of the Navier-Stokes equation that govern the motion of the gas and liquid phases as recently described by us (Zhang et al, 2008). In addition, the continuum-surface-force (CSF) model was invoked to model surface tension acting at the gas-liquid interface.…”

Section: Cfd Modeling Of Shaken 24-srw Microtiter Platesmentioning

confidence: 99%

“…Heath and Kiss (2007) have classified P/V according to its effect on various animal cell-lines. Shear rate (g) and P/V can be directly related by the following equation (Zhang et al, 2008) where m is viscosity…”

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confidence: 99%

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Microwell engineering characterization for mammalian cell culture process development

Barrett

Zhang

et al. 2009

Biotech & Bioengineering

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Experimentation in shaken microplate formats offers a potential platform technology for the rapid evaluation and optimization of cell culture conditions. Provided that cell growth and antibody production kinetics are comparable to those found in currently used shake flask systems then the microwell approach offers the possibility to obtain early process design data more cost effectively and with reduced material requirements. This work describes a detailed engineering characterization of liquid mixing and gas-liquid mass transfer in microwell systems and their impact on suspension cell cultures. For growth of murine hybridoma cells producing IgG1, 24-well plates have been characterized in terms of energy dissipation (P/V) (via Computational Fluid Dynamics, CFD), fluid flow, mixing and oxygen transfer rate as a function of shaking frequency and liquid fill volume. Predicted k(L)a values varied between 1.3 and 29 h(-1); liquid-phase mixing time, quantified using iodine decolorization experiments, varied from 1.7 s to 3.5 h; while the predicted P/V ranged from 5 to 35 W m(-3). CFD simulations of the shear rate predicted hydrodynamic forces will not be detrimental to cells. For hybridoma cultures however, high shaking speeds (>250 rpm) were shown to have a negative impact on cell growth, while a combination of low shaking speed and high well fill volume (120 rpm, 2,000 microL) resulted in oxygen limited conditions. Based on these findings a first engineering comparison of cell culture kinetics in microwell and shake flask formats was made at matched average energy dissipation rates. Cell growth kinetics and antibody titer were found to be similar in 24-well microtiter plates and 250 mL shake flasks. Overall this work has demonstrated that cell culture performed in shaken microwell plates can provide data that is both reproducible and comparable to currently used shake flask systems while offering at least a 30-fold decrease in scale of operation and material requirements. Linked with automation this provides a route towards the high throughput evaluation of robust cell lines under realistic suspension culture conditions.

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Section: Characterization Of Mean Energy Dissipation Ratesmentioning

confidence: 98%

Section: Cfd Modeling Of Shaken 24-srw Microtiter Platesmentioning

confidence: 99%

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Microwell engineering characterization for mammalian cell culture process development

Barrett

Zhang

et al. 2009

Biotech & Bioengineering

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“…Nonetheless, and despite no direct comparison can be made, Zhang et al [49] 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 In an economical view, this allows not only cost reduction with associated equipment, reagents and handling but also it speeds up the development and optimization of bioconversion processes.…”

Section: Time Course Of Typical Bioconversion Runs In Mwp and In Bencmentioning

confidence: 99%

A microwell platform for the scale‐up of a multistep bioconversion to bench‐scale reactors: Sitosterol side‐chain cleavage

Marques¹,

Fernandes²

2010

Biotechnology Journal

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The microwell‐scale approach is widely used for screening purposes and one‐pot biotransformations, but it has seldom been applied to complex whole cell multistep bioconversions, requiring prolonged incubation periods. The present study aims to contribute to filling this gap. The side‐chain cleavage of sitosterol to androstenedione (AD) with Mycobacterium sp. NRRL B‐3805 cells was used as a model system, and focus was given to the screening of suitable bioconversion media with 24‐well microwell plates. Results show that to perform this particular bioconversion growing cells are preferred over resting cells due to higher conversion yields obtained in aqueous medium. The use of resting cells may nevertheless present an interesting approach provided catalytic activity is retained throughout successive runs. Maintaining suitable aeration levels (air flow of 1 mL/min) allowed minimizing the decay of catalytic activity typically observed alongside consecutive bioconversion runs with resting cells. Microwell plates with dedicated oxygen and pH monitoring capabilities proved effective in media development for complex multistep bioconversions using relatively slow‐growing bacteria. Under constant kLa (0.044/s) similar AD production and dissolved oxygen profiles were observed in microwell plates and in a bench‐scale reactor. Selection of a suitable kLa value proved critical, since under lower kLa values scale‐up proved unsuccessful. The same pattern was observed when other scale‐up criteria were evaluated to perform the scale‐up of this particular bioconversion. Results gathered seem to validate the proposed approach “from microwell plate to bench‐scale fermentor”.

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“…Up/down scaling from novel and traditional reactor configurations is still considered a challenging task due to the lack of deterministic equations and detailed fluid mechanics knowledge informing the translation. For example, shaken microwells are commonly used in parallel in bioprocess screening and development (Zhang et al 4 ), but larger scale shaken bioreactors up to 1000 L have been devised at the production level to reduce scaling problematics (Zhang et al 5 ), while in a similar strategy mL scale stirred reactors have entered the market.…”

Section: Introductionmentioning

confidence: 99%

Scalar mixing and strain dynamics methodologies for PIV/LIF measurements of vortex ring flows

Bouremel

Ducci

2017

Physics of Fluids

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Fluid mixing operations are central to possibly all chemical, petrochemical, and pharmaceutical industries, either being related to biphasic blending in polymerisation processes, cell suspension for biopharmaceuticals production and fractionation of complex oil mixtures. This work aims at providing a fundamental understanding of the mixing and stretching dynamics occurring in a reactor in the presence of a vortical structure, and the vortex ring was selected as a flow paradigm of vortices commonly encountered in stirred and shaken reactors in laminar flow conditions. High resolution Laser Induced Fluorescence, LIF, and Particle Imaging Velocimetry, PIV, measurements were carried out to fully resolve the flow dissipative scales and provide a complete data set to fully assess macro-and micro-mixing characteristics.The analysis builds upon the Lamb-Oseen vortex work of Meunier and Villermaux 1 and the Engulfment model of Baldyga and Bourne 2,3 , which are valid for diffusion-free conditions, and a comparison is made between three methodologies to assess mixing characteristics. The first method is the commonly used in macro-mixing studies, and is based on a control area analysis by estimating the variation in time of the concentration standard deviation, while the other two are formulated to provide an insight on local segregation dynamics, by either using an iso-concentration approach or an iso-concentration gradient approach to take into account diffusion.

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Engineering characterisation of a single well from 24-well and 96-well microtitre plates

Cited by 66 publications

References 29 publications

Microwell engineering characterization for mammalian cell culture process development

Microwell engineering characterization for mammalian cell culture process development

A microwell platform for the scale‐up of a multistep bioconversion to bench‐scale reactors: Sitosterol side‐chain cleavage

Scalar mixing and strain dynamics methodologies for PIV/LIF measurements of vortex ring flows

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