• Discovery and Development
Transforming lives by accelerating discovery
• IND enabling preclinical studies
• GLP • AAALAC
• CPCSEA • DSIRS
Keep discovering.
Anthem’s diverse portfolio of services from early in vitro and cell based research to various established animal models, DMPK and toxicology caters to a large number of clients from different industries.
Molecular Biology
• Gene synthesis and cloning
• Expression analysis
• Protein expression
Molecular Biology
Assay Biology
• Enzymatic assays
• Expression assays
• Cell based assays
Assay Biology
ADME/PK
• In vitro DMPK – physicochemical,
distribution, absorption,
metabolism
• In vivo pharmacokinetics
ADME/PK
In vivo Pharmacology
Disease Models:
• Oncology
• Immuno-oncology
• Inflammation
• Auto-immune disorders
• Metabolic disorders
• CNS disorders
In vivo Pharmacology
Toxicology
• Genetic toxicology
• In vivo toxicology services
• In vitro toxicology
• Bio-Analytical Development and
Validation
Toxicology
Microbiology
• In vitro & in vivo evaluation for
Antibacterial and Antifungal
Molecule
• Biofilm evaluation
• AMES test
• Cosmetics/Personal care products
evaluation
• Antibiotic potency evaluation
• Quantitation suspension test /
Time-killassays
• Bacteriophage isolation
Microbiology
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Molecular Biology
Gene synthesis and cloning
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Tailor made vectors through cloning of cDNA, promoter or genomic DNA into any vector for the desired downstream application
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Reporter systems, site-directed mutagenesis, cDNA and genomic DNA libraries
Expression analysis
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mRNA, miRNA and protein isolation from various cell types and tissue samples
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Quantitative analysis of mRNA and miRNA by real time RT-PCR and protein expression analysis performed using Western blotting or ELISA
Protein expression and production
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Bacterial
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Comprehensive services in cloning, expression and purification of peptides and proteins
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Purification of therapeutic proteins, growth hormones, industrial enzymes and viral proteins in bacteria
Yeast -
Bio-process optimization in various yeast expression systems - Saccharomyces cerevisiae, Pichia pastoris
Mammalian -
Transient expression of proteins using plasmid transfections
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Stable expression in engineered cell lines using lentiviral vectors
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High expression levels of heterologous proteins in CHO, Sp2/0 and HEK cell lines
Toxicology
Genetic Toxicology
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Ames test
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Chromosomal aberration test (In vitro & In vivo )
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Micronucleus test (In vitro & In vivo )
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Mouse Lymphoma assay
In vivo toxicology services
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Acute (Six Pack) studies
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Acute Toxicity
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Maximum Tolerated Dose (MTD)
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Dose Range Finding study
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Repeat dose (7 - 90 day) Toxicity studies
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Chronic Toxicity
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Developmental and Reproductive Toxicology (DART) Studies
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Carcinogenicity
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Eye Irritation Test
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Skin Irritation Test
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Skin sensitization (LLNA)
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Eco toxicology services – Studies on Daphnia, fish, earthworm, honeybee, algae
In vitro toxicology
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Ames test
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hERG binding Assay
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Hepatotoxicity
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Cytotoxicity
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Skin Irritation test Episkin
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Skin corrosion test
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3T3 Neutral Red Uptake assay
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In vitro Mammalian Cell Gene Mutation test: L5178YTK+/- Mouse lymphoma
Bio-Analytical Development and Validation
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Pre-formulation development for preclinical studies
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Method development and validation (Formulation & Toxicokinetics)
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Toxicokinetics analysis
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Immunogenicity
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Dose confirmation analysis
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Residue Analysis
Other services
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Phospholipidosis and Steatosis Assay
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Lysosomotropism
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Hemolysis assay
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Transdermal Permeability Testing
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Phototoxicity Testing
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Colony formation cytotoxicity test
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MTT cytotoxicity test
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Filter Diffusion test
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Agar Diffusion test
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Allergenicity Studies
Microbiology
In vitro microbiology
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Anti-infective screening
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Microbial characterization (antibiotic resistance screening)
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Antimicrobial evaluation of compounds by Broth dilution/Micro broth/Agar dilution/Agar well diffusion/disc diffusion method
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MIC determination for antibacterial and antifungal compounds, MIC50, MIC90, MIC with variable elements (serum, albumin, inoculum, temperature effect)
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Kill kinetics (Time Kill), Post-antibiotic effect (PAE)
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Mutation prevention concentration (MPC) and frequency determination
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In vitro Resistance development
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Microbial biofilm susceptibility
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Drug combination study (FIC index, synergism/antagonism /indifference /additive effect)
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Efflux pump inhibitor screen (ethidium bromide accumulation)
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Beta lactamase inhibitor screening
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Hemolytic activity
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S-35 methionine incorporation (for de novo protein synthesis)
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PK-PD correlation
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Bacterial reverse mutation test (Ames test)
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Preservative efficacy test (PET)/ Preservative Challenge Testing
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ISO-11930-2012 Cosmetics
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USP39 – (51) Antimicrobial effectiveness Testing
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Antimicrobial activity
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ASTM: E 2315 – 03 Assessment of Antimicrobial Activity Using a Time-Kill Procedure
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BS EN 1650:2008+A1:2013: Chemical disinfectants and antiseptics
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BS EN 1276:2009: Quantitative suspension test for the evaluation of bactericidal activity
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Sterility test /Total viable count (Product evaluation
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Microbiological Examination
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IS 14648: 2011 Microbiological Examination of Cosmetics and Cosmetic raw material
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Cosmetics/Personal care products evaluation
Bacteriophage
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Isolation of bacteriophages for specific strains
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Characterization – Host range analysis with multiples strains/ One step Growth curve/Burst size/TEM analysis
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Genomic sequence of phages
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Stability studies for bacteriophage formulations
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In vivo efficacy evaluation of bacteriophages
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Available bacterial/fungal strains include multidrug-resistant (MDR) pathogens (ATCC/other)
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Gram Positive strains
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Gram Negative strains
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Yeast & Molds
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Anaerobes and facultative anaerobes
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In vivo microbiology
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Bacterial peritonitis/ Systemic Infection Models/Outcome model – ED50 determination
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Organ burden model
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Thigh – Gram negative/ Gram positive
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Lung
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Vaginal
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Skin/wound
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In vivo Pharmacology
Oncology
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Human xenograft tumor models in immune-compromised mice
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Cell line derived xenograft models
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Orthotopic tumor models
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Metastasis and angiogenesis models
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PK/PD correlation studies
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Biomarker analysis in tumor tissue by IHC/ ELISA/WB/qRT-PCR
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Flow cytometry analysis of tumor tissues
Immuno-oncology models
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Tumor models for Immuno-oncology- Syngeneic tumor models in immunocompetent mice
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Tumor Models: EMT6, CT26, LLC-1, B16F10
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Efficacy of checkpoint inhibitors, Anti-PD1 and Anti-CTLA4, CART-cell therapy
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Tumor microenvironment studies by Flow Cytometry
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Tumor infiltrating lymphocytes (TILs), Tumor associated macrophages
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Cytokine evaluation in Tumors and Serum (IL-6, IP-10, IFNβ)
- Efficacy evaluation combined with Tumor/Blood sampling for Pharmacokinetics, hematology, tumor histology, analysis of proliferation and apoptosis markers (p53, Ki67, TUNEL staining)
Panel of Tumor Models
+ -Origin | Histotype | Cell line |
---|---|---|
Human | Prostate adenocarcinoma | PC3, DU-145, VCaP |
Breast Adenocarcinoma | MDA-MB-231, MCF7, BT-474 | |
Colorectal Adenocarcinoma | HCT-116, HT-29 | |
Ovarian Adenocarcinoma | SK-OV-3, NIH:OVCAR-3 | |
Lung Carcinoma (NSCLC) | A549, NCI-H460, NCIH-1299, NCI-H187 | |
Pancreatic epithelioid carcinoma | PANC-1, MIA PaCa-2 | |
Renal carcinoma | 786-O, A498 | |
Nasopharyngeal carcinoma | FaDu, C666-1, HK1, | |
Glioblastoma | U-87MG | |
Syngeneic (Mouse) |
Lewis lung carcinoma | LLC-1 |
Melanoma | B16F10, B16F0 | |
Colon | CT26 | |
Breast | EMT6 |
Inflammation models
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Inflammatory bowel disease (DSS induced colitis)
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Carrageenan induced edema
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LPS induced sepsis/cytokine release models
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Atopic dermatitis model
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Biomarker analysis
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Inflammatory markers, cytokine analysis in plasma/tissue (TNF-α, IL-6, IL-1), Histopathology and IHC
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Autoimmune disease models
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Rheumatoid Arthritis: Collagen/ Adjuvant-induced Arthritis (CIA) model in DBA1 mice/ Lewis Rats
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Graft Vs. Host disease (GvHD): Allograft GvHD mouse model
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Clinical endpoints: Kaplan-Meier survival analysis, Disease scoring, Histopathology, Cytokine analysis, Flow cytometry
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Imiquimod induced psoriasis
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Immunomodulatory studies: Ex vivo study in Mouse splenocytes
Metabolic diseases and Liver disease models
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Mouse/Rat models of diabetes and obesity
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Genetic models of obesity/diabetes: db/db and ob/ob mice
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Streptozotocin induced diabetes
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High fat induced (HFD) induced diabetes and HFD induced obesity (DIO), dyslipidemia models
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Glucose tolerance studies
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Fatty Liver disease
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CCL4 / Ethanol/ Drug induced hepatotoxicity models
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Clinically relevant endpoint analysis by clinical chemistry, histology, ELISAs, qPCR.
Neuroscience disease models
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Multiple sclerosis: Experimental autoimmune encephalomyelitis (EAE)
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Neurodegenerative disease
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Amyotrophic lateral sclerosis (ALS) in transgenic SOD1G93A mice
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Epilepsy models: Maximum Electroshock (MES) and Pentylenetetrazole (PTZ) induced seizures
Assay Biology
Enzymatic assays
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Anti-oxidants (ORAC/NORAC/SORAC/HORAC)
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NAD/NADH
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Nitric oxide synthesis
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Cox and Lox
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Prostaglandins
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Kinase assays
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Epoxide inhibition assays
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Hydrolase inhibition assays
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Reporter based NF-kb activation assay
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HDAC assay
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Caspase assay
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Cathepsin inhibition assay
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AMPK activation assay
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Protease and DNase assay
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Bead based removal of RNA from DNA sample
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Immunomodulatory effects of ANDA molecules and small molecules
Expression and PTM assays
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mRNA expression using qRT-PCR
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Protein expression through western blot analysis
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AMPK activation assay through ELISA
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Signal transduction assay through western blot analysis
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Aβ-42 estimation
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Cytokine, chemokine and growth factors through ELISA
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Human APP through ELISA
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Target modulation assay for Histone H3 and H4
Cell based assays
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Glucose uptake assay
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Insulin secretion assay
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Differentiation of pre-adipocyte to adipocyte
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ROS assay
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Fatty acid accumulation in HepG2 – Oil O red or Nile red staining
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In vitro NASH model
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In vitro Wilson disease model
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Receptor binding assays
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Migration and scratch assay
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Tube formation (in vitro angiogenesis)
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Cell proliferation and cytotoxicity assay
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ATP synthesis assay
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Mitochondrial aberration – JC-I staining
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Mitotracker analysis
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Phosphorylation
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Early detection of cell apoptosis using Annexin V or caspase assay
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PARP cleavage
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DNA fragmentation
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Cell cycle analysis (PI staining)
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Cell cycle regulator expression study through qRT-PCR or western blot
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Cell based reporter assays (promoter and signal transduction pathway analysis)
ADME/PK
In vitro DMPK
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Physicochemical properties
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Thermokinetic and thermodynamic solubility (Multi-Screen HTS filter plates/ LC- MS/MS method)
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Stability in buffer solutions, simulated gastric fluid and simulated intestinal fluid (Fasted and Fed), in plasma of different species like Mice, Rat, Dog, Monkey and Human
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Log D, log P and pKa determination
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Aqueous solubility and stability at different pH
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Distribution
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Plasma Protein binding determination by Ultrafiltration method and Equilibrium dialysis
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Species: Mice, Rat, Dog, Monkey and Human
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In vitro tissue binding, blood cell partitioning and microsomal binding
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Absorption
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PAMPA (Parallel Artificial Membrane Permeability Assay)
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MDCK-wild type permeability
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MDR1-MDCK permeability for P-gp interaction
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Calcein AM assay for P-gp interaction and inhibition
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Caco-2 permeability – unidirectional and bidirectional permeability
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Metabolism
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Metabolic stability in liver, intestinal microsomes, S9 fractions and hepatocytes
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Metabolite identification (phase I and II) and characterization studies using LC- MS/MS
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Species: mice, rat, dog, monkey and human
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In vivo pharmacokinetics
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Types of PK studies
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Single dose, multiple dose, cross over, cassette dose
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Oral, Intravenous, Subcutaneous, Intradermal, Intramuscular, Intraperitoneal, Per rectal
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rat, mouse, hamsters, guinea pigs, rabbits
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Brain, liver, adipose, skin, muscle, spinal cord
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Cancer models e.g. tumor bearing animals
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Diabetes models e.g. db / db mouse
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Obesity models e.g. Ob/ob mouse, DIO mouse
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Inflammation models e.g. IBD model, arthritis model
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Tissue distribution in efficacy models and PK-PD correlation
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WinNonlin analysis
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Parameters reported: Tmax , Cmax , AUClast , AUC∞, Cl, t1/2 , Vss, MRT and F%
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Method development and validation
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Recovery and linearity studies
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Development of analytical methods in Biological Matrix - plasma, serum, bone marrow, lymph node, urine, feces, bile and tissues
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Sample analysis from both in vivo and in vitro studies
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Facilities
Cell culture suites
- • Fluorescence imaging microscope
- • Multimode plate reader
- • Flow cytometer
- • Cell counter
Vivarium and Toxicology
- • Experiment rooms
- • Procedure rooms
- • Quarantine rooms
- • Surgery room
- • Necropsy room
- • Clinical chemistry and histopathology facility
DMPK – Tox laboratory
- • LC-MS rooms
- • Sample storage rooms
- • Sample preparation rooms
- • Tissue culture lab
- • Test item storage room
- • Archival facility
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