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 PREDIVEZTM 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: PREDEASYTM Kit**

Transporter Cell type ServicesProduct Availability

PREDEASY™Kit**

PREDIVEZ™ Reagent Kit*

Traditional VT Membrane

BCRP (ABCG2)MCF7 ATPase, VT available available available

HEK293 VT – available available

BSEP (ABCB11) Sf9, Hi5, HEK293 VT − − available

MDR1/P-gp (ABCB1)

Human “K” VT − available available

Hi5 ATPase available − −

Sf9 ATPase available − –

HEK293 VT – – available

MRP1 (ABCC1) Sf9 VT − available available

MRP2 (ABCC2)Sf9 ATPase, VT available available available

HEK293 VT – available available

MRP3 (ABCC3)Sf9 ATPase, VT – available available

HEK293 VT − available available

MRP4 (ABCC4) HEK293 VT − − available

MRP5 (ABCC5) HEK293 VT − available available

mouse Bcrp MDCKII VT − − available

mouse BsepSf9 VT − − available

Sf9-HAM ATPase available − −

mouse Mdr1a HEK293 VT − − available

rat Bsep HEK293 VT − − available

rat Mdr1bSf9 ATPase available − –

Sf9-HAM VT − − available

rat Mrp2 HEK293 VT − − available

rat Mrp3 HEK293 VT − − 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

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

Transfected Monolayer Assays

MDR1MDCKII PreadyPortTM 24 & 96 w

LLC-PK1 −

BCRPMDCKII PreadyPortTM 24 & 96 w

LLC-PK1 −rat Mdr1a LLC-PK1 −

mouse Mdr1a LLC-PK1 −rat Bcrp rat Bcrp

mouse Bcrp MDCKII −Double Transfected Monolayer Assays

OATP2B1/BCRP MDCKII PreadyPortTM 24 & 96 wOAT1/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 −CNT1 (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)HEK293 –

CHO 24 & 96 wells

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 −

Monkey & Rodent Transporter Cell Type

Product Availability

PREDICELL™

cyNtcp (Slc10a1) HEK293 −

cyOatp1b1 (Slco1b1) HEK293 −

cyOatp1b3 (Slco1b3) HEK293 −

cyOatp2b1 (Slco2b1) HEK293 −

rNtcp (Slc10a1) CHO −

rOat1 (Slc22a6) CHO −

rOatp1a1 (Slco1a1) HEK293 −

rOatp1a4 (Slco1a4) HEK293 −

rOatp1b2 (Slco1b2) HEK293 −

rOctn2 (Slc22a5) CHO −

Human Transporter Cell TypeProduct

AvailabilityPREDICELL™

OCT2 (SLC22A2)CHO 24 & 96 wells

HEK293 −OCT3 (SLC22A3) HEK293 −

OCTN1 (SLC22A4) CHO 24 & 96 wellsOCTN2 (SLC22A5) CHO 24 & 96 wellsPEPT1 (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 ratSandwich 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 24- and 96-well plates available available available

Rat 24- and 96-well plates available available available

Monkey 24- and 96-well plates available available available

Dog 24-well plates available − −Multiple Species Customized

In collaboration with Ascendance Biotechnology, Inc.

RENAL PROXIMAL TUBULE CELL MONOLAYER SERVICES: human, rat and mouseThe Renal Proximal Tubule cell (PTC) monolayer model is a unique in vitro approach for investigating renal drug handling. Utilizing freshly-isolated primary human or rodent 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 & reabsoption – Transporter inhibition – DDI – Clinically Relevant Biomarker Production.

In partnership with Newcastle University, UK.

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.

StudiesStandard Setups

Basic Studies at one concentration

Extended Studies at multiple concentrations

Passive permeability studies: Uni- and bidirectional permeability available available

Substrate assessment: Specific transporter studies

available Mdr1a

available Mdr1a

Inhibitor assessment: Drug-Drug Interaction (DDI) studies

available Mdr1a

available Mdr1a

DDI - IC50 determination for quinidine inhibition, positive control: PSC833

available Mdr1a

available Mdr1a

EXCIPIENTS & ABSORPTION (Caco-2 model)MDR1/P-gp Inhibition Assessment • BCRP Inhibition Assessment • Effect on Passive Permeability

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.

human 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.

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

PRICES ARE AVAILABLE UPON REQUEST AT SALES@SOLVO.COM