02115nas a2200205 4500008004100000022001400041245015200055210006900207260007300276300001000349490000700359520127600366653003001642653001701672653002701689653002701716100001701743700001901760856013001779 2008 eng d a0167-701200aSimple whole cell based high throughput screening protocol using Mycobacterium bovis BCG for inhibitors against dormant and active tubercle bacilli0 aSimple whole cell based high throughput screening protocol using aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcAPR a62-680 v733 a

This study aimed at developing a whole cell based high throughput screening protocol to identify inhibitors against both active and dormant tubercle bacilli. A respiratory type of nitrate reductase (NarGHJI), which was induced during dormancy, could reflect the viability of dormant bacilli of Mycobacterium bovis BCG in microplate adopted model of in vitro dormancy. Correlation between reduction in viability and nitrate reductase activity was seen clearly when dormant stage inhibitor metronidazole and itaconic anhydride were applied in this in vitro microplate model. Active replicating stage could also be monitored in the same assay by measuring the A(620) of the culture. MIC values of 0.08, 0.075, 0.3 and 3.0 mu g/ml, determined through monitoring A(620) in this assay for rifampin, isoniazid, streptomycin and ethambutol respectively, were well in agreement with previously reported by BACTEC and Bio-Siv assays. S/N ratio and Z' factor for the assay were 8.5 and 0.81 respectively which indicated the robustness of the protocol. Altogether the assay provides an easy, inexpensive, rapid, robust and high content screening too] to search novel antitubercular molecules against both active and dormant bacilli. (C) 2008 Elsevier B.V. All rights reserved.

10ahigh throughput screening10atuberculosis10aWayne's dormancy model10awhole cell based assay1 aKhan, Arshad1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/simple-whole-cell-based-high-throughput-screening-protocol-using-mycobacterium-bovis-bcg-001811nas a2200229 4500008004100000022001400041245009700055210006900152260006700221300001400288490000700302520091800309653003001227653003201257653003001289653004001319653004301359100002001402700001601422700001901438856012401457 2012 eng d a8756-793800aIn situ pH maintenance for mammalian cell cultures in shake flasks and tissue culture flasks0 aIn situ pH maintenance for mammalian cell cultures in shake flas a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcNOV a1605-16100 v283 a

pH in animal cell cultures decreases due to production of metabolites like lactate. pH control via measurement and base addition is not easily possible in small-scale culture formats like tissue-culture flasks and shake flasks. A hydrogel-based system is reported for in situ pH maintenance without pH measurement in such formats, and is demonstrated to maintain pH between 6.8 and 7.2 for a suspension CHO cell line in CD CHO medium and between 7.3 and 7.5 for adherent A549 cells in DMEM:F12 containing 10% FBS. This system for pH maintenance, along with our previous report of hydrogels for controlled nutrient delivery in shake flasks can allow shake flasks to better mimic bioreactor-based fed batch operation for initial screening during cell line and process development for recombinant protein production in mammalian cells. (C) 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012

10ahigh throughput screening10apH control for cell culture10apH control in shake flask10apH control in tissue culture flasks10asmall scale platforms for cell culture1 aPradhan, Ketaki1 aPant, Tejal1 aGadgil, Mugdha uhttps://library.ncl.res.in/content/situ-ph-maintenance-mammalian-cell-cultures-shake-flasks-and-tissue-culture-flasks-002344nas a2200229 4500008004100000022001400041245016900055210006900224260006700293300001200360490000700372520139200379653002001771653002001791653002101811653003001832653002401862653003901886653003901925100001901964856013101983 2015 eng d a0268-257500aDevelopment of a mathematical model for animal cell culture without pH control and its application for evaluation of clone screening outcomes in shake flask culture0 aDevelopment of a mathematical model for animal cell culture with a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcJAN a166-1750 v903 a

BACKGROUNDCurrently used mathematical models for animal cell growth and metabolism cannot simulate changing culture pH and its effect on metabolism, and hence cannot accurately model cultures lacking pH control. Animal cell cultures are however widely cultured at a small scale without pH control, e.g. for clone screening during cell line development. A mathematical model for animal cell growth without pH control is developed and applied to design optimal conditions for clone screening without pH control. RESULTSThe mathematical model successfully simulates published data for suspension CHO culture in shake flasks. It is further used to simulate clonal heterogeneity and predict relative clone performance in different culture conditions. Predictions of correlation coefficients between clone titers in batch and fed batch screening conditions and pH controlled fed batch agree with published values. Fed batch mode is predicted to give better screening outcomes than batch, but only for clones able to consume lactate. Simulations reveal that culture duration of the screening assay has a large effect on the screening outcome and optimal durations differ based on the ability or inability of clones to consume lactate. CONCLUSIONThis study presents a tool for optimal design of screening assay for clone selection without pH control. (c) 2014 Society of Chemical Industry

10aclone screening10aclone selection10aculture duration10ahigh throughput screening10alactate consumption10amathematical model with pH effects10apH control for animal cell culture1 aGadgil, Mugdha uhttps://library.ncl.res.in/content/development-mathematical-model-animal-cell-culture-without-ph-control-and-its-application-0