The scope of Biology services at Eurofins Advinus encompasses comprehensive in vitro and in vivo profiling of molecules for their activity in various biochemical and cell-based systems and animal models.

Our facilities are designed to meet the most stringent requirements of both national and international regulatory agencies. Biology activities at Eurofins Advinus are carried out in strict accordance with guidelines from regulatory bodies such as the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Institutional Animal Ethics Committee (IAEC), Institutional Biosafety Committee (IBSC) and Atomic Energy Regulatory Board (AERB).

In vitro Biology

Eurofins Advinus provides customized / fit-for-purpose solutions for in vitro research for a broad range of target classes and phenotypic readouts. Our services include:

1. Small molecule screening

• Activity assays for enzymes, nature of binding and enzyme kinetics.
• Formats: Fluorescence, Absorbance, Fluorescence Polarisation, Luminescence, FRET, TR-FRET, AlphaScreen, NanoBret
• Through-put: 96 and 384
• Binding studies by SPR/BIACORE and FACS.
• Functional assays: cAMP, β-arrestin, IP-1 modulation
• Binding studies by SPR, Fluorescence Polarisation,
• Ternary complex formation assays.
• Protein degradation assays (WB/JESS)
• Protein ubiquitination

2. Large molecule screening

• Affinity determination: Binding studies by SPR (BIACORE 8K) and FACS for monoclonals and engineered antibodies.

3. Therapeutic area specific cell-based functional assays

A. Autoimmunity and Inflammation
• Immune cell activation assays
• T/B/NK cell activation assays using mitogens
• Macrophage differentiation assays
• M1/M2 macrophage differentiation from monocytes
• DC differentiation from monocytes
• MDSC induction
• T cell differentiation and polarization assays
• Treg/Th1/Th2/Th17 differentiation
• T cell cytotoxicity assay
• Cytotoxic T lymphocyte assay (OT-1 system)
• Primary immune cell profiling by flow cytometry
• Immune cell suppressive assay
• MDSC/Treg/Tumor-T suppressive assay
• Antibody mediated immunological assay
• Immune cell populations and subsets – frequency, activation, and differentiation
• Cytokine and chemokine analysis
• Mixed Lymphocyte reaction assay
• Cytokine release assay by ELISA and intracellular cytokine assay by Flowcytometry

B. Oncology
• Cytotoxicity assays (growth kinetics/ Incucyte platform)
• Angiogenesis Assay
• LDH, Propidium Iodide and CFSE labelled Cell Cycle Based Assays
• Apoptosis Assay
• Cell motility, invasion
• Chemotaxis assay
• 2D-clonogenic assay
• Cell migration assays
• Cell invasion assay
• Phosphorylation studies and detection by WB or JESS
• Cell proliferation assays (growth rate, doubling time, viability)
• Cell death assays (apoptosis, necrosis, autophagy)
• Cell cycle assays (cell cycle phases, mitosis)

C. Immuno-oncology assays
• Immune cell profiling in tumor infiltrating leukocyte (TIL) and other tissues (spleen, blood, lymph node, liver, etc.) profiling:
• Immune cell population analysis (T, B, NK, DC, Myeloid, Macrophage, Granulocyte, CD4+T, CD8+T, Treg, Th1/2/17, naïve/memory/effector T, g/mMDSC, M1/M2, etc.).
• Functional and intracellular marker analysis (PD-1, PD-L1/2, CTLA-4, TIM-3, LAG-3, OX40, 4-1BB, ICOS, Ki-67, IL-2, IL-17, IFN-γ, TNFα, Granzyme B, etc.).
• Immune and cancer cell coculture assays

D. Metabolic Diseases
• Glucose uptake; Glycogen synthesis; Glycolysis
• Cells: Primary cultures and cell lines such as 3T3L1, C2C12, L6, HepG2 Glucose uptake assay in primary hepatocytes and adipocytes
• Glycogen synthesis in primary hepatocytes
• Endogenous glucose production in primary hepatocytes
• Immuno-fluorescence studies to monitor translocation of proteins in cell
• Glucose-dependent Insulin secretion in rodent islets
• Glucagon secretion in rodent islets
• Fatty acid oxidation in skeletal muscle cells [ex vivo; soleus muscles]
• Lipolysis in primary adipocytes
• GLP-1 secretion in entero-endocrine cells
• Rat primary brown adipose tissue (BAT) cultures to study metabolic parameters/biomarkers
• Rat primary cardiomyocyte culture to study metabolic parameters/biomarkers

E. Ex-vivo studies: Immune cell profiling
• Cell subset analysis: CD4, CD8, MDSC, M1/M2, DC cell, NK cell, B cell
• Exhaustion marker analysis: PD1, TIM3, LAG3, OX40, 4-1BB
• Activation marker analysis: CD25, CD69, CD62L, B220, MHC Class II, CD80, CD86
• Cytokine release by ELISA and flowcytometry
• Lymphoid tissue and peripheral blood analysis
• Immune cell populations and subsets – frequency, activation, and differentiation
• Cytokine and chemokine analysis
• IPT and TDAR

F. Biomarker analysis

• Soluble biomarker analysis by ELISA
• Cell-based biomarker by FACS and Western Blot
• Gene expression profiling by RT-PCR
• Tumor infiltrating lymphocytes (tils) analysis on human and murine tumor tissues by flow cytometry

Plasmid DNA and Molecular Cloning
• Transformation, competent cell preparation and bacterial cultures
• Blunt end and sticky end based cloning methods
• Insertion of oligonucleotides, introduction of site directed mutagenesis
• Sequence and ligation free cloning methods
• Systems: DH5α, Stbl3, JM109
Transcript Expression
• RNA extraction from cell lines, fresh, frozen and FFPE samples
• SYBR green and Taqman methods for quantitative real time PCR
• Copy number and relative expression analysis
In vitro transcription (IVT)
• ARCA and CleanCap based mRNA synthesis
• Cy5 labelling of mRNA
Protein Expression and quantification
• Native and SDS PAGE
• Western blotting/JESS
• ELISA/FACS/Biacore
• Co-immunoprecipitation

Cell Engineering
• Plasmid/Lentiviral/Adenoviral/BACMAM vector based stable and transient transfections
• Electroporation and lipid nanoparticle-based transfections.
• Reporter gene-based promoter assays
• IRES based correlated expression of multiple proteins
• siRNA and shRNA-based gene knock-down
• HA/FLAG/reporter gene tagged protein expression
• Organelle specific protein expression (eg. Golgi, ER, Nucleus, membrane)
Cellular imaging
• Immunofluorescence staining methods
• Live and fixed cell imaging

• Mice immunisation
• Titer determination
• Fusion for hybridoma generation
• Limiting dilution for single cell cloning
• ELISA and FACS based screening of monoclonals

TAT: 4 months

In vivo Pharmacology

  • State-of-the-art in vivo research infrastructure for lower species:
  • Mice, including Immunosuppressed
  • Rats
  • Rabbits
  • Approved for breeding and experiments by Indian regulatory authorities such as the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA)
  • Institutional Animal Ethics Committee (IAEC) is in place according to CPCSEA guidelines.
  • Import licenses in place from Indian Government to enable procurement of animals from the USA, Europe and Japan.
  • The in vivo Pharmacology team has extensive experience in setting up novel in vivo pharmacodynamics (PD) and efficacy models, tailor-made to meet client needs with aggressive timelines. These models are supported by strong expertise in “PK-PD correlation” ex vivo assays, histopathology, and PD marker studies.

List of Animal Models