TheL-Asparagine/L-Glutamine/Ammoniatestkitisanovelmethodforthespecific,convenient,costeffectiveandrapidmeasurementandanalysisofL-asparagine,L-glutamineandammoniaasacrylamideprecursorsinthefoodindustry,orascellculturemedia/supernatantcomponents,orinothermaterials.
ExtendedcofactorsstABIlity.Dissolvedcofactorsstablefor>1yearat4oC.
Oncolyticmeaslesvirusesproducedatdifferentscalesunderserum‐freeconditions.
Weiss,K.,Gerstenberger,J.,Salzig,D.,Mühlebach,M.D.,Cichutek,K.,Pörtner,R.&Czermak,P.(2015).EngineeringinLifeSciences,15(4),425-436.
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Measlesvirus(MV)withattenuatedpathogenicityhaspotentialasoncolyticagent.However,theclinicaltranslationofthistherapyconcepthasonemajorhurdle:theproductionofsufficientamountsofinfectiousoncolyticMVparticles.ThecurrentstudydescribesoncolyticMVproductioninVerocellsgrownonmicrocarrierusingserum-freemedium.Theimpactofthenumberofharvests,cellconcentrationatinfection(CCI),multiplicityofinfection(MOI),andtemperatureonMVproductionwasdeterminedindifferentproductionscales/systems(staticT-flasks,dynamicspinner,andbioreactorsystem)andmodes(batch,repeated-batch,andperfusion).Cellgrowth,metabolic,andproductionkineticswereanalyzed.ItwasfoundthatthenumberofharvestshadthestrongestpositiveimpactonMVyieldineachproductionscale,andthathightemperaturesaffectedMVyieldadversely.ModerateMVtiterswereproducedinT-andspinnerflasksat37°C(~107 TCID50 mL-1,whereTCID50istissuecultureinfectivedoses50%),butstirredtankreactor(STR)MVproductionat37°Cyieldedupto10 000-foldlowerMVtiters.Incontrast,atlowertemperatures(32°C,27°C),1.4×107 TCID50 mL-1,wereachievedintheSTR.VariationsinMOIandCCIhadalmostnoinfluenceonMVproductionyield.ThecurrentstudyimprovesoncolyticMVproductionprocessunderstandingandidentifiesprocessbottlenecksforlarge-scaleproduction.
UV-methodforthedeterminationofL-Asparagine,L-Glutamine
andAmmoniainpotatoes,foodstuffsandcellculturemedia
Principle:
(glutaminase)
(1)L-Glutamine+H2O→L-glutamate+NH4+
(microbialglutamatedehydrogenase)
(2)NH4++2-oxoglutarate+NADPH→L-glutamate+NADP++H2O
(asparaginase)
(3)L-Asparagine+H2O→L-aspartate+NH4+
Kitsize: *50assaysofeach
* Thenumberofmanualtestsperkitcanbedoubledifallvolumesarehalved.
ThiscanbereADIlyaccommodatedusingtheMegaQuantTM Wave
Spectrophotometer(D-MQWAVE).
Method: Spectrophotometricat340nm
Reactiontime: ~20min
Detectionlimit: 0.50mg/L(L-asparagine)
0.54mg/L(L-glutamine)
0.06mg/L(ammonia)
Applicationexamples:
Potatoes,potatoproducts,vegetables,cerealsandothermaterials
(e.g.BIOLOGicalcultures,samples,etc.)
Methodrecognition: Novelmethod
Advantages
- Veryrapidreactionduetouseofuninhibitedglutamatedehydrogenase
- AllenzymessuppliedasstabilisedsUSPensions
- Onlykitavailable
- Verycosteffective
- Allreagentsstablefor>2yearsafterpreparation
- Mega-Calc™softwaretoolisavailablefromourwebsiteforhassle-freerawdataprocessing
- Standardincluded
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