VESICULAR TRANSPORT ASSAYS - Solvo Biotech · VESICULAR TRANSPORT ASSAYS The Vesicular Transport (VT) Assay is an effective tool for inhibition (IC 50 or K i value determination)

VESICULAR TRANSPORT ASSAYSThe Vesicular Transport (VT) Assay is an effective tool for inhibition (IC50 or Ki value determination) as well as substrate identification for efflux transporters. The assay utilizes isolated membranes from cells overexpressing the transporter of interest. The assay is performed in the presence and absence of ATP, and transporter-free negative control membranes are available. Product kit for IC50 determination: Traditional VT Membranes and PREDIVEZ

TM Reagent Kit*

ATPase ASSAYSThe ATPase Assay is an in vitro assay designed to indicate the nature of the interaction between a compound and an efflux (ATP-binding cassette; ABC) transporter. ABC transporters mediate the transport of substrates against a concentration gradient using energy derived from ATP hydrolysis, which is stimulated by transported substrates, and can be measured using a colorimetric method. Using the ATPase activity, both activation and inhibition of transporters can be investigated. Deliverables: EC50 and IC50 values. Product kit: PREDEASY

TM Kit**

Transporter Cell type ServicesProduct Availability

PREDIVEZ™ Reagent Kit*

PREDEASY™Kit**

Traditional VT Membrane

BCRP (ABCG2)MCF7 ATPase, VT available available available

HEK293 VT available – availablecyBcrp HEK293 VT − − available

mouse Bcrp MDCKII VT − − availableBSEP (ABCB11) Sf9, Hi5, HEK293 VT − − available

cyBsep HEK293 VT – − availabledogBsep HEK293 VT – − available

mouse BsepSf9 VT – − available

Sf9-HAM ATPase – available −rat Bsep HEK293 VT – − available

MDR1/P-gp (ABCB1)

Human “K” VT available − availableHi5 ATPase − available −Sf9 ATPase − available –

HEK293 VT – – availablecyMdr1 HEK293 VT – − available

mouse Mdr1a HEK293 VT – − available

rat Mdr1bSf9 ATPase – available −

Sf9-HAM VT – – availableMRP1 (ABCC1) Sf9 VT available − available

MRP2 (ABCC2)Sf9 ATPase, VT available available available

HEK293 VT available − availablerat Mrp2 HEK293 VT − − available

MRP3 (ABCC3)Sf9 ATPase, VT available − available

HEK293 VT available − availablerat Mrp3 HEK293 VT − − available

MRP4 (ABCC4) HEK293 VT − − availableMRP5 (ABCC5) HEK293 VT available − available

*: Membranes and appropriate Control Membranes are not included | **: Membranes are included

Transporter Cell Type Product AvailabilityCaco-2 Monolayer Assays

Caco-2 or C2BBe1 (Caco-2 clone)CacoReadyTM 24 & 96 wCacoGobletTM 24 w

BCRP Caco-2 or C2BBe1-BCRP-KO −MDR1 Caco-2 or C2BBe1-MDR1-KO −MRP2 Caco-2 or C2BBe1-MRP2-KO −

Transfected Monolayer Assays

BCRPMDCKII PreadyPortTM 24 & 96 w

LLC-PK1 −

MDR1MDCKII PreadyPortTM 24 & 96 w

LLC-PK1 −cyBcrp MDCKII −

mouse Bcrp MDCKII −mouse Mdr1a LLC-PK1 −

rat Bcrp MDCKIIrat Mdr1a LLC-PK1 −

Double Transfected Monolayer Assays OATP2B1/BCRP MDCKII PreadyPortTM 24 & 96 w

OAT1/BCRP MDCKII −OAT3/BCRP MDCKII −

OCT2/MATE1 MDCKII −OCT2/MATE2-K MDCKII −

Transporter-specific C2BBe1 knockouts were developed by MilliporeSigma. PreadyPort kits are available in transfected, parental, or in mixed format including both transfected and parental cells. The cell type for PreadyPort kits is MDCKII.

UPTAKE TRANSPORTER ASSAYSThe Uptake (SLC) Transporter Assay utilizes stably transfected cells for inhibition (IC50 or Ki value determination), as well as substrate studies (accumulation studies, KM and Vmax determination).

MONOLAYER ASSAYSMonolayer Assays are performed using polarized cell monolayers expressing one or more efflux or uptake transporters. They are suitable for inhibition studies (e.g IC50 determination), or for investigation of time- and concentration-dependent permeability of small molecules (substrate assessments). Transporter knockout (KO) Caco-2 cells are a good alternative for determining the potential impact of specific transporters in drug absorption and disposition without dependence on chemical inhibitors.

Human Transporter Cell TypeProduct

AvailabilityPREDICELL™

ASBT (SLC10A2) HEK293 24 & 96 wellsCNT1 (SLC28A1) MDCKII −CNT2 (SLC28A2) MDCKII −CNT3 (SLC28A3) MDCKII −ENT1 (SLC29A1) MDCKII −ENT2 (SLC29A2) MDCKII −ENT4 (SLC29A4) HEK293 −HPT1 (SLC47A1) MDCKII −

MATE1 (SLC47A1)CHO –

MDCKII 24 & 96 wellsMATE2-K (SLC47A2) MDCKII 24 & 96 wells

NTCP (SLC10A1)CHO 24 & 96 wells

HEK293 –

OAT1 (SLC22A6)CHO 24 & 96 wells

HEK293 –OAT2 v1 (SLC22A7) HEK293 −

OAT3 (SLC22A8)MDCKII 24 & 96 wellsHEK293 −

OATP1B1 (SLCO1B1) CHO, HEK293 24 & 96 wellsOATP1B3 (SLCO1B3) CHO, HEK293 24 & 96 wellsOATP1A2 (SLCO1A2) HEK293 24 & 96 wellsOATP2A1 (SLCO2A1) CHO −OATP2B1 (SLCO2B1) MDCKII 24 & 96 wells

OCT1 (SLC22A1)CHO 24 & 96 wells

HEK293 −

OCT2 (SLC22A2)CHO 24 & 96 wells

HEK293 −

Monkey & Rodent Transporter Cell Type

Product Availability

PREDICELL™cyNtcp (Slc10a1) HEK293 −

cyOatp1b1 (Slco1b1) HEK293 −cyOatp1b3 (Slco1b3) HEK293 −cyOatp2b1 (Slco2b1) HEK293 −

rAsbt (Slc11a2) HEK293 −rNtcp (Slc10a1) CHO −rOat1 (Slc22a6) CHO −

rOatp1a1 (Slco1a1) HEK293 −rOatp1a4 (Slco1a4) HEK293 −rOatp1b2 (Slco1b2) HEK293 −rOctn2 (Slc22a5) CHO −

rOst α/β (Slc51a and b) HEK293 –

Human Transporter Cell TypeProduct

AvailabilityPREDICELL™

OCT3 (SLC22A3) HEK293 −OCTN1 (SLC22A4) CHO 24 & 96 wellsOCTN2 (SLC22A5) CHO 24 & 96 wells

OSTα/β (SLC51A and B) HEK293 –PEPT1 (SLC15A1) CHO −PEPT2 (SLC15A2) CHO −SGLT1 (SLC5A1) HEK293 −SGLT2 (SLC5A2) HEK293 −

URAT1 (SLC22A12) CHO, MDCKII −

HEPATOCYTE UPTAKE: human and rat*Hepatocyte Uptake assays utilize cryopreserved primary hepatocytes to investigate temperature-, time- and concentration-dependent accumulation of small molecules. Filter plate and oil-spin seperation methods are available.

Standard Uptake – Km/Vmax Determination – Drug-drug interaction studies

* Other species are available upon request

SANDWICH-CULTURED HEPATOCYTE SERVICES using B-CLEAR®: human and rat*Sandwich cultured hepatocyte (SCH) assays utilize primary hepatocytes grown between layers of collagen, which allows retention of physiologicial morphology and function, including expression and localization of basolateral and canalicular transporters and formation of canalicular bile pockets (analogous to bile canaliculi in vivo). Using patented B-CLEAR® technology, the tight junctions surrounding the bile pockets can be disrupted, enabling determination of biliary efflux and clearance, and biliary transporter interactions.

Biliary excretion – Hepatic transporter inhibition – Hepatic DDI assessments

*B-CLEAR® licensed from Qualyst Transporter Solutions | Other species are available upon request

HepatoPac® micropatterned hepatocyte co-cultures: human, monkey, rat and dogHepatoPac® is a powerful in vitro tool for studying hepatic transport, metabolism, and toxicity. It utilizes micropatterned co-cultures (MPCCs) of primary hepatocytes and stromal cells, simulating the micro-scale in vivo architecture of the liver, and allowing the retention of physiological transporter and metabolic enzyme expression and activity over several weeks in culture. Potential applications include xeno- and endobiotic transport studies, metabolite identification and analysis, induction studies, and mechanistic and predictive toxicology studies, with long-term multiple dosing. MPCC Services as well as HepatoPac® Kits are available.

Species Package MetID Metabolic Stability ToxicityHuman, rat, monkey 24- and 96-well plates available available available

Dog 24-well plates available − −Multiple Species Customized

In collaboration with Ascendance Biotechnology.

PROXTOX-HTS: rapid and accurate in vitro high-throughput prediction of nephrotoxicity in humansProxTox-HTS is the first high throughput in vitro screening method developed to predict nephrotoxicity in humans with high accuracy. ProxTox-HTS utilizes freshly-isolated primary human renal proximal tubule (PTC) or HK-2 cells in a 384-well format, combined with powerful high content imaging analysis and machine learning. ProxTox-HTS has been pre-validated using 44 chemically diverse compounds with well-characterized effects on human kidneys. The test balanced accuracies using ProxTox-HTS are 82% (primary PTCs) and 89% (HK-2 cells), respectively.

www.proxtox.com | In partnership with IBN, A*STAR, Singapore.

RENAL PROXIMAL TUBULE CELL MONOLAYER SERVICES: human, rat, mouse, dog, and monkeyThe Renal Proximal Tubule cell (PTC) monolayer model is a unique in vitro approach for investigating renal drug handling. Utilizing freshly-isolated primary human or preclinical animal proximal tubule cells, physiological expression and function of transporters, metabolizing enzymes, and signaling proteins is retained. Suitable for many applications, including xenobiotic transport (substrate and inhibition assays), transport of endogenous molecules (eg. creatinine, urate, lactate, or phosphate), receptor-mediated endocytosis, investigation of signaling pathways, cross-species differences, and predictive or mechanistic toxicity studies utilizing clinically-relevant biomarkers.

Renal excretion & reabsorption – Transporter inhibition – DDI – Clinically Relevant Biomarker Production.

In partnership with Newcells Biotech.

ABSORPTION & EXCIPIENT (Caco-2 model)Caco-2 models can be utilized for a variety of different measurements related to absorption and drug-interactions on the intestinal level. The assay has been characterized for studying permeability (BCS Biowaiver compatible), transport of endogenous molecules (e.g. sugars), drug-drug effects of inhibitors or excipients (MDR1/P-gp and BCRP inhibition), as well as identifying involvement of individual transporter in knock-out Caco-2 cells (MDR1-KO, BCRP-KO and MRP2-KO).

MOLECULAR IMAGINGSmall-animal positron emission tomography (PET) is an imaging technique which allows measuring the tissue distribution and pharmacokinetics of radiolabelled molecules non-invasively in living animals (e.g. mice, rats, rabbits). For PET imaging positron-emitting radionuclides (11C, 18F, 68Ga, 124I) need to be incorporated into the molecule of interest. PET data combined with magnetic resonance imaging (MRI) data provide detailed anatomical information about the tissue distribution of radio-labelled molecules.

microPET & microMRI / Transporter-specific PET

• Small-animal (e.g. mice, rats, rabbits) positron emission tomography (PET) to measure the tissue distribution and pharmacokinetics of radiolabelled molecules non-invasively

• Combined with magnetic resonance imaging (MRI) data to provide detailed anatomical information about the tissue distribution

• Customized PET tracers: carbon-11(11C), fluorine-18(18F), gallium-68(68Ga) or iodine-124(124I)

• Assessment of radiotracer metabolism

• In vivo drug distribution studies

• In vivo drug-drug interaction studies in animals

In collaboration with the Austrian Institute of Technology | Clinical PET studies are also available.

LC-MS/MS QUANTIFICATION OF TRANSPORTER PROTEINS

SOLVO’s study design is flexible. The main experimental parameters, including probe substrate, reference inhibitor, and number of concentrations or replicates can be changed according to individual requirements. ©2016 SOLVO Biotechnology 111 Huntington Avenue, Boston, MA 02199. V 2.

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18human BCRP, BSEP, MCT2, MDR1/P-gp, MRP2, NTCP, OCT1, OCT2, OAT1, OAT2, OAT3, OATP1B1, OATP1B3, OATP2B1, PEPT1 & Na+/K+ ATPase

mouse Abcb4, Abcc4, Bcrp, Bsep, Mdr1a, Mdr1b, Mrp2, Mrp3, Ntcp, Oatp1a1, Oatp1a4, Abcg8, Abcg5 & Na+/K+ ATPase

In collaboration with Bertin Pharma.

CHEMICALSNMQ - MDR1/P-gp Substrate

NMQ (N-methyl-quinidine) is a hydrophilic monoquaternary drug

Quantities available: 5, 10, 50, 100, 500 mg, and 1 g

PRICES ARE AVAILABLE UPON REQUEST AT [email protected]

RAT BRAIN ENDOTHELIAL CELL MONOLAYER ASSAYSThe Rat Brain Endothelial Cell (RBEC) monolayer assay studies the brain penetration of compounds by evaluating their vectorial transport across an endothelial cell monolayer, analogous to in vivo brain capilaries. This system is a co-culture of three primary cell types: endothelial cells, pericytes and astrocytes, which ensures barrier formation and expression of key enzymes, including transporters.