Development Of An Immunomagnetic Method Based On IgG1 Monoclonal .
Development of anImmunomagnetic method basedon IgG1 Monoclonal antibodiesfor Cryptosporidium and GiardiaPhilip Scandizzo, Graham Vesey,Mark Gauci, David Baer, JamesSmith, Duncan Veal*
WATER CONCENTRATEConcentrated WaterSampleAdd MagneticBeadsGIARDIA CYSTSCRYPTOSPORIDIUM OOCYSTSMAGNETIC BEADSMAGNETIC DEBRISOTHER DEBRIS
PRECLEARApply magneticfieldGIARDIACRYPTOSPORIDIUM OOCYSTSMAGNETIC BEADSMAGNETIC DEBRISOTHER DEBRISMAGNET
Addition of ConjugatedMagnetic BeadsAdd antibody conjugatedmagnetic beadsGIARDIA CYSTSCRYPTOSPORIDIUM OOCYSTSMAGNETIC BEADSOTHER DEBRISMAGNETIC BEAD CONJUGATED TO GIARDIA MABMAGNETIC BEAD CONJUGATED TO CRYPTO MAB
IMS OF CRYPTOSPORIDIUM& GIARDIAApply magneticfieldMAGNETGIARDIA CYSTSCRYPTOSPORIDIUM OOCYSTSOTHER DEBRISMAGNETIC BEAD CONJUGATED TO GIARDIA MABMAGNETIC BEAD CONJUGATED TO CRYPTO MAB
Separation of (oo)cysts fromdebrisRemove SupernatantRe-suspendMAGNETMAGNETGIARDIA CYSTSCRYPTOSPORIDIUM OOCYSTSMAGNETIC BEADSMAGNETIC BEAD CONJUGATED TO GIARDIA MABMAGNETIC BEAD CONJUGATED TO CRYPTO MAB
Acid DissociationAdd 0.1M HClGIARDIA CYSTSCRYPTOSPORIDIUM OOCYSTSMAGNETIC BEAD CONJUGATED TO GIARDIA MABMAGNETIC BEAD CONJUGATED TO CRYPTO MAB
Acid DissociationMAGNET
IdentificationFLUORESCENTMAB STAININGMABS G203 Cry 104
Analysis by flow cytometryMABS Cry 104 G203FACSLaserDetectorCYTOMETERNon dissociated bead(oo)cyst complex
Recovery of oocysts from Wimalee Lagoon concentratedwater sample and PBS using the Immucell IMS Kit.PBSWimalee LagoonRecovery (%) 36 4.633 6.4DAPI Positive (%) 25 4.021 2.6Time Taken microscopy 17 2.1(min)20 1.5
Recovery of oocysts from Wimalee Lagoon concentratedwater sample and PBS using the Dynal IMS KitPBSRecovery (%)DAPI Positive (%)Time Takenmicroscopy (min)WimaleeLagoon47 6.5 36 3.428 4.9 25 3.319 2.6 20 2.0
Beads and antibodiesBeadsSize Supplier( m)Dynal IgG 4.5BiomagSpherotechDynal IgM1Dynal ,Costbeads /ml 145Perspective 6Diagnostics4-4.5 Spherotech, 204.5 145Dynal ,Cryptosporidium andGiardia-specific mAbCRY 104 (Mac Uni)OW50 (Uni Arizona);Giardia-specific mAbG203 (Mac Uni)CRY 104 (MacquarieUniversity)CRY 104 (MacUni)CRY 26 (Mac Uni)
Recovery of oocysts from Wimalee Lagoon using 0W50coated anti-IgG Dynal beads and CRY 26-coated antiIgM Dynal beads.Recovery (%)Time Taken FACS(min)Time Takenmicroscopy (min)0W50-coated antiIgG Dynal beads71 ( 12.8)5.2 ( 0.4)CRY 26-coated anti-IgMDynal beads.56 ( 17.5)5.9 ( 0.6)8.8 ( 1.2)7.7 ( 0.4)
Comparison of recovery of oocysts from Wimalee Lagoonperforming IMS with CRY 104-coated Spherotech ,Biomag and Dynal -anti-IgG beads.Recovery (%)Time Taken FACS(min)Time Takenmicroscopy (min)Biomag 91 ( 3.1)11.4 ( 2.3)Dynal 94 ( 3.4)10.1 ( 1.4)Spherotech 79 ( 7.5)12.7 ( 1.2)12 ( 1.0)13 ( 1.5)12.3 ( 1.5)
Recovery of oocysts from different water types using CRY104-coated Dynal beads.WaterSampleOriginalsamplevolume (L)Recovery(%)Time Takenmicroscopy(min)78.3 ( 5.5)TimeTakenFACS(min)10.3 ( 2.2)4.5 ( 1.0)WimaleeLagoonDrinkingWaterRiverWater1090 ( 3)10010092.7 ( 5.8)5.9 ( 2.1)14.3 ( 2.5)14.3 ( 2.5)19.3 ( 5)
Recovery of Giardia cysts from different water types usingG203-coated Dynal anti-IgG beadsWater TypeRecovery(%)Time TakenFACS (min)WimaleeLagoonDrinkingWaterRiver Water86.3 ( 5.5)5.1 ( 0.5)Time Takenmicroscopy(min)3.8 ( 0.2)87 ( 7.9)5.5 ( 0.5)5.4 ( 0.2)78.3 ( 11.9)5.6 ( 0.7)4.4 ( 0.5)
Dynal IgG 4.5 Dynal , 145 CRY 104 (Mac Uni) OW50 (Uni Arizona); Giardia-specific mAb G203 (Mac Uni) Biomag 1 Perspective Diagnostics 6 CRY 104 (Macquarie University) Spherotech 4-4.5 Spherotech, 20 CRY 104 (MacUni) . Recovery of oocysts from different water types using CRY
EPA Test Method 1: EPA Test Method 2 EPA Test Method 3A. EPA Test Method 4 . Method 3A Oxygen & Carbon Dioxide . EPA Test Method 3A. Method 6C SO. 2. EPA Test Method 6C . Method 7E NOx . EPA Test Method 7E. Method 10 CO . EPA Test Method 10 . Method 25A Hydrocarbons (THC) EPA Test Method 25A. Method 30B Mercury (sorbent trap) EPA Test Method .
The Problem with Root Cause Analysis Method A Method B Method C Method G Method E Method H Method J Method F Method D Method I No‐one can agree on "what is a root cause." Everyone says they do "root cause analysis,"yet everyone is doing something different!
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G2 F30 instrument (FEI Company, Hillsboro, OR, USA) was used to capture TEM images at operated 120 kV. For TEM measurements, 10 μL of nanoparticle solution was dropped and waited for 10 min. FEI Nova NanoSEM 430 microscope (FEI, Eindhoven, Netherlands) was used to get SEM images. Bacteria concentrations were adjusted using
Magnet. Read the manual prior to using the device. It is essential that the operator have theoretical knowledge as well as lab-scale practical experience with Immunomagnetic Separation (IMS). Please pay special attention to the Notes, Warnings and Limitations provided throughout the User
A practical approach of method development under QbD principles - Screening - Optimization - Robustness study, Design of Experiments - Transfer & Verification . Agilent Solutions for QbD Method Development ISET Intelligent System Emulation Technology RA Remote Advisor Harmonized Qualification ACE Method Development System & Method Scouting SW . Agilent Method Development Systems 15 1352 .
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