In vitro data provides invaluable information on the compounds chances to succeed at the clinical phase. GVK BIO offers a panel of validated in vitro pharmacological assays that cover a broad range of targets. Apart from ascertaining the potency of the NCE on target, assays are also available for selectivity screening and profiling. These assays are available either as stand-alone or as a screening platform.
GVK Bio’s in vitro pharmacology panel of services include the following:
 Assay Development
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Molecular Biology Techniques
- Molecular (drug target) cloning
- Transfection (transient or/and stable) & expression optimization studies in insect & mammalian cells
- Recombinant cell membrane preparation for binding assays
Assay standardization and Validation
- Protein expression and assay development plan
- Assay validation based on established practices
- Identification of physical & statistical parameters critical for assay validation
- Standard operating procedures (SOP) for assay
- Assay run with a set of reference compounds with rank order potency and ascertained for reproducible data
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Receptor biology
The receptor biology lab at GVK BIO has capabilities to perform receptor binding and functional assays. We have in house molecular biology capabilities to generate recombinant cell lines for receptor binding and functional studies.
Receptor binding assays:
Radioligand binding assays are developed for several therapeutic targets such as GPCRs, ion channels, transporters etc. using membranes from recombinant cells as well as native tissues.
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Muscarinic
• M1 (Human)
• M2 (Human)
• M3 (Human)
• M4 (Human)
• M5 (Human)
Opoid
• µ (Human)
• KAPPA (Human)
• DELTA (Human)
Dopamine
• D1 (Human)
• D2L (Human)
• D4.4 (Human)
• D5 (Human)
Serotonin
• 5HT1A (Human)
• 5HT2A (Human)
• 5HT2C (Human)
• 5HT3 (Human)
• 5HT7 (Human)
Histamine
• H1 (Human)
• H2 (Human)
Adenosine
• A1 (Human)
• A2A (Human)
Norepinephrine Transporter-Net (Human)
Dopamine Transporter-Dat (Human) -3
Serotonin Transporter-5htt (Human)
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GPCR functional assays:
As an extension to our receptor binding assay platform, we have developed functional assays in house to screen the GPCR targets. Ligand binding to GPCR promotes G protein coupling initiating signal transduction pathways which trigger a series of cellular responses.
GVKBIO has capabilities for measuring GPCR functional responses by developing cAMP assays for Gs coupled receptors and Ca2+ or IP3/PIP2 measurements for Gq coupled receptors. GVKBIO also has capabilities for performing Guanine nucleotide binding assay ([35S] GTPγS assay) for Gi coupled receptors.
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Enzyme assays
Enzymes are increasingly recognized as potential drug targets in drug discovery. GVKBIO offers assays for several enzyme classes using different readouts like fluorescence, luminescence, time resolved fluorescence and radiometry. The following are a few of our enzymes in our database which are validated in-house. GVKBIO has the capabilities to develop and validate client specific customized enzyme assays.
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Kinases
• PKA (cAMP-dependent kinase)
• PKC (a, b, g, d)
• p38(a)-MAPK
• CDK-1/CyclinB
• Src-Tyrosine Kinase
Proteases
• Caspase-3 & Caspase-8
• Cathepsin-B & Cathepsin-L
• Matrix Metalloproteases {MMP-1, 2, 3, 7, 12 & 13}
• Dipeptidyl Protease-IV (DPP-4)
• Angiotensin Converting Enzyme (ACE)
• TNF- a Converting Enzyme (TACE)
Phosphatases
• PTP-1B
Monoamine Oxidase
• MAO-A
• MAO-B
Cyclooxygenases
• Cyclooxygenase-1 (COX-1)
• Cyclooxygenase-2 (COX-2)
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Specialized Cellular assays
Cellular assays are one of the prime screening technologies used in pre-clinical drug discovery process, starting from primary screening to in vitro toxicology. GVK BIO’s focus is on developing relevant, accurate and informative cell-based systems that can simulate in vivo conditions.
The following assays are standardized at GVK BIO which represents in house capabilities and infrastructure for conducting cell based assays. |
Cytokine assays using PBMCs
• TNF-α secretion
• IL-1β secretion
• IL-6 secretion
Cell Proliferation (Thymidine incorporation/ MTT/ Alamar blue)
• PDGF stimulated
• Serum stimulated
• PHA stimulated
Cytotoxicity
• Cell viability
Adhesion assays
• ICAM-1
• VCAM-1
Cycloxygenases using Whole blood assays
• COX-1
• COX-2
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