Description
Non-PEGylateddoxorubicinliposomecomesinathree-vialkitandhasalipidcompositionandliposomesizesimilartocommercialMyocet®.Doxorubicindrugcannotbepre-loadedtotheliposomesinthesamewaythatispre-loadedintothePEGylatedliposomes.ThematrixliposomeinPEGylatedliposomeishydrogenatedsoyPCwhichisasaturatedlipidandcreatesatightpackedliposomesuitableforpre-loADIngandholdingthedruginsidetheliposomes.ThematrixlipidinthisformulationisL-⍺-phosphatidylcholinewhichisanunsaturatedlipid.Itmakesaloosepackedmembraneandtherefore,itcannotholdthedrugforalongtime.Thedoxorubicindrugshouldbeloadedintotheliposomesmomentsbeforeuse.
Thekitcomesinthreevials.Thefirstvialcontainsaveryconcentratedsolutionofliposomes.TheliposomesaremadeincitratebufferatpH4.ThesecondvialiscomposedofsodiumcarbonateatpH11.4.Addingvial2tovial1makesapHgradientaroundthelipidmembrane.DuetothedifferenceinthepHinsideandoutsideoftheliposomes,therewillbeaconstantfluxofprotonfromtheinsidetotheoutsideoftheliposomes.Duetothisflux,doxorubicinmoleculesmoveintheoppositedirectionfromtheoutsidetotheinsideoftheliposomes.Duringtheremoteloadingprocess,duetopHgradient,thedoxorubicindrugisabletofreelypassintotheliposomeswhereitisboundbyhydrogenandnanocrystalsofdoxorubicincitratesalttoformlargerdoxorubicincompounds.Asthenewdoxorubicincomplexesareformed,theycometogethertoformoneofthethreedifferentshapesinsidetheliposomes.Alongsingularbandthatstretchesthemembrane,aclosedcircularbandoranopenedU-shapedband.ThecircularandU-shapedcomplexesarethemostcommonones.DoxorubicinliposomesthatareformedbyremoteloadingusingpHgradientshouldbeusedimmediately.
PEGylatedformulationofdoxorubicinliposomesisalsoavailable.Formoreinformationseehere.
FormulationInformation
Doxosome™Kit-DoxorubicinLiposomes(Non-PEGylated)
| 3-VialKitforDoxosome™(Non-PEGylated) | Specification |
|---|---|
| Vial1 | PreformedliposomescomposedofHSPCandcholesterol(55:45molarratio) |
| Vial2 | SodiumcarbonateformakingapHgradient |
| Vial3 | DoxorubicinHClin0.9%saline |
| Instructions:Add0.4mloftheliposome(vial1)to0.6mlofthesodiumcarbonatesolution(vial2).Thenadd10mgofthedoxorubicinin4mlofthe0.9%salinesolution(vial3)tothemix,heatat55-60°Cfor7min,andshakeitfor10s.Thefinalconcentrationofdoxorubicinintheliposomesis2mg/mlandthetotalvolumeofthereagentis5ml.Over95%ofthedrugisloadedtotheliposomes.Thedrugtolipidw/wratiois0.27:1. | |
| LipidCompositionforVial1 | Concentration(mg/ml) | Concentration(mM) | MolarRatioPercentage |
|---|---|---|---|
| Total | 100mg/ml | 166.8mM | 100 |
| L-alpha-Phosphatidylcholine | 71.3 | 92.6 | 55 |
| Cholesterol | 28.7 | 74.2 | 45 |
| BufferandLiposomeSizeforVial1 | Specification |
|---|---|
| Buffer | CitrateBuffer |
| pH | 4(in300mlbuffer) |
| LiposomeSize | 180nm |
| Vial2 | Specification |
|---|---|
| SodiumCarbonate | 166mM |
| pH | 11.4 |
| Vial3 | Specification |
|---|---|
| DoxorubicinHCl | Solutionin0.9%Saline |
LiposomeParticleCalculator
Doxorubicinliposomesareunilamellarandsizedto100nm.Themolarconcentrationofliposomeis166.8mM.Byhaving liposomediameter(nm)andlipidconcentration(µM),youcancalculatethetotalnumberofthelipidsinoneliposomeandthenumberoftheliposomesinonemilliliteroftheliposomesolution.Tousethecalculatorclick here.
Appearance
Doxosome™isaredtranslucentliquidmadeofnanosizeunilamellarliposomes.Usually,duetothesmallsizeofliposomes,nosettlingwilloccurinthebottomofthevial.Theliposomesarepackagedinanambervial.
EducationalVideo
Ordering/ShippingInformation
- Allliposomebasedformulationsareshippedonblueiceat4°Cininsulatedpackagesusingovernightshippingorinternationalexpressshipping.
- LiposomesshouldNEVERbefrozen.Icecrystalsthatforminthelipidmembranecanrupturethemembrane,changethesizeoftheliposomesandcausetheencapsulateddrugtoleakout.Liposomesinliquidformshouldalwaysbekeptintherefrigerator.
- ClientswhoorderfromoutsideoftheUnitedStatesofAmericaareresponsIBLefortheirgovernmentimporttaxesandcustomspaperwork.EncapsulaNanoSciencesisNOTresponsibleforimportationfeestocountriesoutsideoftheUnitedStatesofAmerica.
- WestronglyencouragetheclientsinJapan,Korea,TaiwanandChinatoorderviaadistributor.Toughcustomsclearanceregulationsinthesecountrieswillcausedelayincustomclearanceoftheseperishableformulationsifordereddirectlythroughus.Distributorscaneasilyclearthepackagesfromcustoms.Toseethelistofthedistributorsclickhere.
- ClientsorderingfromuniversitiesandresearchinstitutesinAustraliashouldkeepinmindthattheliposomeformulationsaremadefromsyntheticmaterialandtheformulationsdonotrequirea“permittoimportquarantinematerial”.LiposomesareNOTBIOLOGicalproducts.
- Ifyouwouldlikeyourinstitute’sFedExorDHLaccounttobechargedforshipping,thenpleaseprovidetheaccountnumberatthetimeofordering.
- EncapsulaNanoScienceshasnocontroloverdelaysduetoinclementweatherorcustomsclearancedelays.YouwillreceiveaFedExorDHLtrackingnumberonceyourorderisconfirmed.ContactFedExorDHLinadvanceandmakesurethatthepaperworkforcustomsisdoneontime. AllsubsequentshippinginquiriesshouldbedirectedtoFederalExpressorDHL.
StorageandShelfLife
Storage
Doxosome™productsshouldalwaysbestoredatinthedarkat4°C,exceptwhenbroughttoroomtemperatureforbriefperiodspriortoanimaldosing.DONOTFREEZE.IfthesUSPensionisfrozen,theencapsulateddrugcanbereleasedfromtheliposomesthuslimitingitseffectiveness.Inaddition,thesizeoftheliposomeswillalsochangeuponfreezingandthawing.
ShelfLife
Doxosome™ismadeondailybasis.Thebatchthatisshippedismanufacturedonthesameday.Itisadvisedtousetheproductswithin4monthsofthemanufacturingdate.
Referencesandbackgroundreading
1.HermansonGT.Bioconjugatetechniques.Academicpress;2013Jul25.
2.TorchilinV,WeissigV,editors.Liposomes:apracticalapproach.OxfordUniversityPress;2003Jun5.
3.GrabarekZ,GergelyJ.Zero-lengthcrosslinkingprocedurewiththeuseofactiveesters.Analyticalbiochemistry.1990Feb15;185(1):131-5.
4.YanL,CraytonSH,ThawaniJP,AmirshaghaghiA,TsourkasA,ChengZ.ApH‐ResponsiveDrug‐DeliveryPlatformBasedonGlycolChitosan–CoatedLiposomes.Small.2015Oct1;11(37):4870-4.
5. Silva-LópezEI,EdensLE,BardenAO,KellerDJ,BrozikJA.ConditionsforliposomeadsorptionandbilayerformationonBSApassivatedsolidsupports.Chemistryandphysicsoflipids.2014Oct31;183:91-9.
6.HazraM,SinghSK,andRayS.SurfaceModificationofLiposomalVaccinesbyPeptideConjugation.JournalofPharmaSciTech,2011;1(1):41-47.
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我想用酶联免疫斑点（ELISPOT）技术测定NK细胞活性，但不知道哪里能买到试剂盒，特向大家求助，谢谢。
尤其是获得SCI文献引用的产品，
绝对ok的
请问一般的实验室可以进行ELIspot吗？（我可以买到现成的试剂盒，资金这个应该没有问题。）ELIspot的难点在哪里？多谢大虾们的指点！这个对我很重要，多谢！！！
