Evotec

Expression of target protein in bacteria (exemplified by but not limited to AVI101, BL21 and derivatives and DH5a) in shake flasks or fermentor with expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be transformation.

Evotec is uniquely qualified in delivering its customers small and large molecule library management needs through over a decade of being the world’s leading sample management operation. Evotec expanded its operational presence in 2013 and again in 2015 by adding capacity and capabilities in Branford, Connecticut and Toulouse, France. These facilities are strategically positioned to support the strong presence of Evotec’s drug discovery services along the east coast of the U.S. and across Europe. Our service offering includes: Cost effective compound identification, selection, and procurement High-throughput compound analysis Multi-format plating and reformatting (including nano volume) Inert or dry atmosphere and low temperature storage and processing Fast order fulfilment and worldwide delivery Multi-site disaster recovery and business continuity “In-sourcing” of Evotec sample management staff to client sites Access to compound collections of some of its partners through an open innovation strategy Evotec has a broad range of sample management expertise across many sectors including large Pharma, biotechs, foundations and governmental organisations; which results in a deep understanding of its customers’ needs and deadlines. Each client has a dedicated Project Manager, single point of contact ensuring an open line of communication is always maintained.

Evotec offers a broad expertise in providing recombinant proteins. Working in partnership with our clients we offer customised solutions, bespoke and modular workflows, flexible production scale and access to a unique suite of integrated in-house technologies, to deliver projects in a time-efficient and cost-effective manner. Strong communication driven by scientist-to-scientist interaction. A core team of Bioinformaticians, Structural Biologists and Protein Scientists design protein constructs and production routes tailored to their final use, whether it be assay, biophysics, activity assays, structural biology or animal studies. Steps in optimisation of construct design include: Bioinformatics analysis including homology modelling and secondary structure prediction using state-of-the-art software Identify optimal tagging strategies, domain boundaries and mutagenesis Choice of expression host and desired cell localisation e.g. secreted, periplasmic Custom vector design Multiple expression systems: E. coli Yeast Baculovirus infected insect cells Mammalian cells Cell-free

Expression of target protein in yeast in shake flasks or fermentor with expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be transformation.

Expression of target protein in mammalian cells, in shake flasks, wave bag or bioreactor (exemplified by but not limited to HEK 293 and CHO cells). Expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be a stable cell line or transfection. Transfection reagents are not included in pricing and will be charged separately.

Evotec offers a broad expertise in providing recombinant proteins. Working in partnership with our clients we offer customised solutions, bespoke and modular workflows, flexible production scale and access to a unique suite of integrated in-house technologies, to deliver projects in a time-efficient and cost-effective manner. Strong communication driven by scientist-to-scientist interaction. A core team of Bioinformaticians, Structural Biologists and Protein Scientists design protein constructs and production routes tailored to their final use, whether it be assay, biophysics, activity assays, structural biology or animal studies.

Steps in optimisation of construct design include:
Bioinformatics analysis including homology modelling and secondary structure prediction using state-of-the-art software
Identify optimal tagging strategies, domain boundaries and mutagenesis
Choice of expression host and desired cell localisation e.g. secreted, periplasmic
Custom vector design

Multiple expression systems:
E. coli
Yeast
Baculovirus infected insect cells
Mammalian cells
Cell-free
Comprehensive purification platform:
Multiple cell lysis options
Protein refolding screen and scale-up by dilution/dialysis
Chromatography including, affinity, ion exchange, hydrophobic interaction, size exclusion and native
Endotoxin removal and confirmation by LAL assay
Typical QC includes SDS-PAGE, western blot, Mass Spec, Dynamic Light Scattering, High-Throughput analytical/fluorescence size exclusion, biophysics and activity assays, SEC-MALLS

Membrane Proteins
Working with an optimised process for high-throughput screening of expression and solubilisation conditions and large scale protein production, we deliver quality isolated target protein, amenable for assays, biophysics and structural biology, in fast turnaround times.
Scouting and sample optimisation:
Fluorescence based SEC screening
Automated microscale affinity purification
Thermofluor based stability assay on membrane bound and solubilised sample
Radioligand binding assay on membrane bound and solubilised sample
LCP-FRAP and in situ screening through Diamond light source, UK

Large scale membrane protein production:
Heterologous expression in mammalian, insect, yeast and bacterial systems
Production of Virus Like Particles (VLPs) and lipoproteins
Enriched membrane preparation and purification
Pharmacological and biophysical analysis for characterisation of proteins
Crystallisation in lipid cubic phase (LCP), bicelles and vapour diffusion

Labelling strategies include isotope labels for NMR and mass spectroscopy standards, SeMet enriched proteins for macromolecular crystallography and biotinylation for assay and SPR.

Evotec also offers labelling of biologically relevant molecules and reagents for use in ELISA, FACS, FCS, plate readers and other fluorescence based detection systems. Our service helps clients to improve their research results in all areas of biological and pharmacological assays, as well as clinical diagnostics, analytics and agrochemicals.

Our expertise covers the labelling of the following species:
Peptides and chemokines (as enzyme substrates, GPCR ligands, etc.)
Proteins including antibodies
Site-specific mutagenesis of proteins (for site-specific labelling)
Glycoproteins
Carbohydrates

For labelling purposes we use the optimally suited dyes and tags for your application, such as:
Commercially available fluorescent dyes and the in-licensed MR121 dye
Non-fluorescent markers and probes like affinity tags, biotin, reporter enzymes, etc.

High-Throughput parallelised protein production

For fast and efficient production of proteins we offer a high-throughput platform for protein expression scouting and purification. Robust statistical models are employed to triage constructs. Informed decision-making to maximise throughput and minimise waste. The platform has been successfully employed over more than 1,500 unique constructs. The platform is suitable for E. coli, baculovirus infected insect cells and mammalian cell lines.
Standardised high-throughput protein production work flow:
Expression Scouting of constructs in parallel
In-house statistical model for review and prioritisation
Large Scale Expression (1-3 liters)
Parallel Purification and QC
Blocks of 96 constructs from gene-to-protein within 2-3 months, ready for multiple downstream experiments

Evotec offers a comprehensive purification platform: Multiple cell lysis options Protein refolding screen and scale-up by dilution/dialysis Chromatography including, affinity, ion exchange, hydrophobic interaction, size exclusion and native Endotoxin removal and confirmation by LAL assay Typical QC includes SDS-PAGE, western blot, Mass Spec, Dynamic Light Scattering, High-Throughput analytical/fluorescence size exclusion, biophysics and activity assays, SEC-MALLS

Purification of secreted or intracellular protein from cells derived from E.coli, yeast, insect, mammalian cells. Lysis of cells (exemplified by but not limited to sonication or microfluidization) followed by a single-column or batch purification using technologies exemplified by but not limited to His-Tag, GST-tag, ion exchange, heparin, hydroxyapatite, or hydrophobic interaction chromatography. The soluble and insoluble cell lysis fractions, column flow-through and elution peak fractions are characterized by SDS-PAGE and western blot (to affinity tag or protein). Analysis can include Dynamic Light Scattering (DLS), analytical size exclusion (aSEC), mass spectrometry (LC-MS), SDS-PAGE, N-terminal sequencing, HSQC (NMR), deglycosylation and LC-MS/MS as needed.

Evotec offers a broad expertise in providing recombinant proteins. Working in partnership with our clients we offer customised solutions, bespoke and modular workflows, flexible production scale and access to a unique suite of integrated in-house technologies, to deliver projects in a time-efficient and cost-effective manner. Strong communication driven by scientist-to-scientist interaction. A core team of Bioinformaticians, Structural Biologists and Protein Scientists design protein constructs and production routes tailored to their final use, whether it be assay, biophysics, activity assays, structural biology or animal studies. Steps in optimisation of construct design include: Bioinformatics analysis including homology modelling and secondary structure prediction using state-of-the-art software Identify optimal tagging strategies, domain boundaries and mutagenesis Choice of expression host and desired cell localisation e.g. secreted, periplasmic Custom vector design Multiple expression systems: E. coli Yeast Baculovirus infected insect cells Mammalian cells Cell-free Comprehensive purification platform: Multiple cell lysis options Protein refolding screen and scale-up by dilution/dialysis Chromatography including, affinity, ion exchange, hydrophobic interaction, size exclusion and native Endotoxin removal and confirmation by LAL assay Typical QC includes SDS-PAGE, western blot, Mass Spec, Dynamic Light Scattering, High-Throughput analytical/fluorescence size exclusion, biophysics and activity assays, SEC-MALLS Membrane Proteins Working with an optimised process for high-throughput screening of expression and solubilisation conditions and large scale protein production, we deliver quality isolated target protein, amenable for assays, biophysics and structural biology, in fast turnaround times. Scouting and sample optimisation: Fluorescence based SEC screening Automated microscale affinity purification Thermofluor based stability assay on membrane bound and solubilised sample Radioligand binding assay on membrane bound and solubilised sample LCP-FRAP and in situ screening through Diamond light source, UK Large scale membrane protein production: Heterologous expression in mammalian, insect, yeast and bacterial systems Production of Virus Like Particles (VLPs) and lipoproteins Enriched membrane preparation and purification Pharmacological and biophysical analysis for characterisation of proteins Crystallisation in lipid cubic phase (LCP), bicelles and vapour diffusion Labelling strategies include isotope labels for NMR and mass spectroscopy standards, SeMet enriched proteins for macromolecular crystallography and biotinylation for assay and SPR. Evotec also offers labelling of biologically relevant molecules and reagents for use in ELISA, FACS, FCS, plate readers and other fluorescence based detection systems. Our service helps clients to improve their research results in all areas of biological and pharmacological assays, as well as clinical diagnostics, analytics and agrochemicals. Our expertise covers the labelling of the following species: Peptides and chemokines (as enzyme substrates, GPCR ligands, etc.) Proteins including antibodies Site-specific mutagenesis of proteins (for site-specific labelling) Glycoproteins Carbohydrates For labelling purposes we use the optimally suited dyes and tags for your application, such as: Commercially available fluorescent dyes and the in-licensed MR121 dye Non-fluorescent markers and probes like affinity tags, biotin, reporter enzymes, etc. High-Throughput parallelised protein production For fast and efficient production of proteins we offer a high-throughput platform for protein expression scouting and purification. Robust statistical models are employed to triage constructs. Informed decision-making to maximise throughput and minimise waste. The platform has been successfully employed over more than 1,500 unique constructs. The platform is suitable for E. coli, baculovirus infected insect cells and mammalian cell lines. Standardised high-throughput protein production work flow: Expression Scouting of constructs in parallel In-house statistical model for review and prioritisation Large Scale Expression (1-3 liters) Parallel Purification and QC Blocks of 96 constructs from gene-to-protein within 2-3 months, ready for multiple downstream experiments

Expression of target protein in bacteria (exemplified by but not limited to AVI101, BL21 and derivatives and DH5a) in shake flasks or fermentor with expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be transformation.

Expression of target protein in insect cells, in shake flasks, wave bag or bioreactor (exemplified by but not limited to High5, Sf9, Sf21, T. ni or S2 cells). Expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be a high titer stock or BIICs.

Expression of target protein in yeast in shake flasks or fermentor with expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be transformation.

Expression of target protein in mammalian cells, in shake flasks, wave bag or bioreactor (exemplified by but not limited to HEK 293 and CHO cells). Expression confirmation by SDS-PAGE and Western blot (to affinity tag or protein). Starting point will be a stable cell line or transfection. Transfection reagents are not included in pricing and will be charged separately.

Evotec offers stable cell line generation services through a customised offering tailored to our partner`s needs. Cell lines are produced from recombinant cells which have been either generated by stable transfection or viral transduction. Working from highly stable clones, a broad range of target expression levels and co-expression of multiple sub-units is available. All cell lines are screened using immunofluorescence labelling, Western blot, or RT-PCR, and further validated in cell-based assays to confirm the functional expression of the target protein. Cell line generation at Evotec Cell line generation capabilities include: Custom expression strategies and vector design Constitutive or inducible target expression Target or reporter gene expression Polycistronic or independent co-expression of auxiliary factors Multiple subunits and resistance markers

Evotec has a long history in high-throughput screening utilising multiple compound collections and has run more than 320 cell based, micro-organism based or biochemical HTS campaigns. Leveraging this expertise, a state-of-the-art screening platform, a strong experience in assay development and miniaturisation (>475 assays developed), and an excellent disease biology knowledge, Evotec selects the most suitable screening strategy for your target.

Evotec typically uses a three-part incoming Quality Control (“QC”) process comprised of weight, solubility and LC-MS purity and identity to ensure that all collections start with high-quality compounds. Systems include a Waters 8-channel LC-MS system, with digital UV, ELSD and an LCT Premier mass spectrometer, and two Waters LCT Premier XE uPLC systems featuring positive and negative ESI and UV and ELS detection. Evotec also utilises a Xevo G2 Q-TOF system for quantitative determination and high resolution of compounds in DMSO solution.

Evotec has extensive experience in the identification, selection and procurement of small molecule compounds possessing very specific (including commercial, physicochemical or bioactivity-related) properties. Evotec has the capability to apply compound identification tools as required by the client.
Once compounds are identified for purchase, our Project Management group consults with selected suppliers to negotiate terms on behalf of our client requirements, e.g. price, purity and format. Evotec purchases tens of thousands of compounds on a regular basis for a number of clients; therefore, the client can benefit from our vendor relations and volume discounts.

In the pharmaceutical industry, early liability assessment of drug candidates is an important element to decrease the high attrition rate in the drug discovery and development process. One of the key challenges is the balance between drug efficacy and potential adverse effects at the earliest possible point in time for de-risking cost-intensive activities especially during late-stage clinical development.

To meet this demand, Evotec has established an industrialised high-throughput ADME (absorption, distribution, metabolism and excretion) profiling service for routine compound screening against a panel of the most critical safety pharmacology targets. In support of discovery chemistry programmes, externalisation of safety pharmacology profiling is providing a unique business opportunity for Evotec’s customers to reserve internal capacity for core research activities, expanding their ADME assay portfolio and to mitigate risk. Besides these benefits, high-throughput ADME profiling is a time- and cost-efficient process consistently delivering high-quality data.

At Evotec, we perform both unbiased biomarker discovery and hypothesis validation within a disease-relevant context. As a result, we develop and make use of disease-mimicking animal models to validate biomarkers within the context of a living organism. Most importantly, where possible, we work directly with patient material to ensure clinical translatability.

Relying on our strong drug discovery expertise, we aim to identify for our partners:
•Pharmacodynamic, efficacy and target engagement biomarkers

•Stratification biomarkers to support the identification of a target patient population

•Alternate tissue or pathway read-outs (surrogate biomarkers)

•Safety biomarkers to monitor toxicity

OUR OFFERING COMPRISES:

• Tailor-made workflows enabling either proteome profiling for quantitative analysis across hundreds of samples, or deep proteome profiling for detection of more than 10,000 proteins from cells or tissues
  In-depth, quantitative protein biomarker discovery in body fluid samples such as plasma and CSF
• Full capabilities to develop targeted mass spectrometry assays based on multiple reaction monitoring (MRM) and parallel reaction monitoring (PRM) for high-throughput verification and validation of investigational biomarkers
• In vitro and in vivo targeted metabolomics enabling various pre-clinical and clinical translational biomarker applications
• Leading expertise in statistics and bioinformatics for the identification and validation of univariate and multivariate markers in the context of relevant disease models or patient data

Successful drug discovery requires a deep understanding of biological context coupled with knowledge and experience of the property space required to deliver safe, efficacious drugs; DMPK continues to be a key player in this success since the major sources of attrition – clinical efficacy and safety – are clearly linked to exposure in key tissues either on- or off-target.

Evotec and its wholly owned subsidiary Cyprotex offer a comprehensive and flexible range of in vitro and in vivo DMPK/ADME studies designed to cover the majority of activities in the hit-to-lead and lead optimisation phase through to candidate selection. We also offer in vitro metabolite profiling, structural elucidation and DDI (drug-drug interaction) studies for IND and NDA submissions during pre-clinical development and clinical stage projects.

There is realisation that evaluating toxicology earlier in a project reduces the risk of late-stage failure. As a consequence, human relevant cell-based models are being introduced in drug discovery to address this need. Cyprotex is leading the science in this field and have significant expertise in the latest techniques such as HCS, MEA and 3D cell-based models.

The scientific expertise and processes can be accessed as fee-for-services or as part of partially or fully integrated programmes. DMPK and ADME-Tox support will be provided to our partners in a variety of ways tailored to suit their specific project requirements.

At Evotec, we perform both unbiased biomarker discovery and hypothesis validation within a disease-relevant context. As a result, we develop and make use of disease-mimicking animal models to validate biomarkers within the context of a living organism. Most importantly, where possible, we work directly with patient material to ensure clinical translatability.

Relying on our strong drug discovery expertise, we aim to identify for our partners:
•Pharmacodynamic, efficacy and target engagement biomarkers

•Stratification biomarkers to support the identification of a target patient population

•Alternate tissue or pathway read-outs (surrogate biomarkers)

•Safety biomarkers to monitor toxicity

OUR OFFERING COMPRISES:

• Tailor-made workflows enabling either proteome profiling for quantitative analysis across hundreds of samples, or deep proteome profiling for detection of more than 10,000 proteins from cells or tissues
  In-depth, quantitative protein biomarker discovery in body fluid samples such as plasma and CSF
• Full capabilities to develop targeted mass spectrometry assays based on multiple reaction monitoring (MRM) and parallel reaction monitoring (PRM) for high-throughput verification and validation of investigational biomarkers
• In vitro and in vivo targeted metabolomics enabling various pre-clinical and clinical translational biomarker applications
• Leading expertise in statistics and bioinformatics for the identification and validation of univariate and multivariate markers in the context of relevant disease models or patient data

At Evotec, we perform both unbiased biomarker discovery and hypothesis validation within a disease-relevant context. As a result, we develop and make use of disease-mimicking animal models to validate biomarkers within the context of a living organism. Most importantly, where possible, we work directly with patient material to ensure clinical translatability.

Relying on our strong drug discovery expertise, we aim to identify for our partners:

• Pharmacodynamic, efficacy and target engagement biomarkers
• Stratification biomarkers to support the identification of a target patient population
• Alternate tissue or pathway read-outs (surrogate biomarkers)
• Safety biomarkers to monitor toxicity

OUR OFFERING COMPRISES:

• Tailor-made workflows enabling either proteome profiling for quantitative analysis across hundreds of samples, or deep proteome profiling for detection of more than 10,000 proteins from cells or tissues
  In-depth, quantitative protein biomarker discovery in body fluid samples such as plasma and CSF
• Full capabilities to develop targeted mass spectrometry assays based on multiple reaction monitoring (MRM) and parallel reaction monitoring (PRM) for high-throughput verification and validation of investigational biomarkers
• In vitro and in vivo targeted metabolomics enabling various pre-clinical and clinical translational biomarker applications
• Leading expertise in statistics and bioinformatics for the identification and validation of univariate and multivariate markers in the context of relevant disease models or patient data

Pharmacology Services

Drug Discovery & Development Services

Cells and Tissues Services

Hit Identification Services

Drug Discovery Services

Product Development, Testing, and Packaging Services

Analytical Chemistry Services