Bioassay
Every great biological model developed for functional target and biomarker validation or screening the effects of novel therapeutics, be it tissue-based, cellular or biofluid, needs a suite of bioassays to measure and quantify biological change.
To support our cellular biology services, our skilled scientific team deliver quantitative data from a comprehensive range of assay endpoints, including mediator/cytokine release, automated Western, changes in gene expression, immunocytochemistry (ICC) and cellular imaging.
ELISA – specific analysis of key biomarkers
A robust and proven technology, enzyme linked immunosorbent assays (ELISAs) deliver quantitative antibody-based measurement of any target for which there is a sensitive and specific binding reagent, such as an antibody or aptamer.
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Verification and execution of commercial ELISAs across a range of matrices (plasma, serum, urine, cell lysates, cell supernatants)
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Assay transfer (custom assays or commercial kits), verification, and execution
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Multiple platforms: chromogenic, fluorescent, luminescent
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Cytokine and biomarker analysis
Automated Westerns – quantitative & precise targeted protein analysis
Immunoblotting is a widely used analytical technique to detect and quantify specific proteins in a complex mixture, such as cell lysates or tissue homogenates, by exploiting the specificity of antibody-antigen interactions.
Our team can analyse samples by traditional SDS-PAGE & Western blotting, but in recent years has adopted the Jess as our platform of choice. Jess is an advanced automated platform from Biotechne that delivers accurate, reproducible and quantitative data. This, teamed with our experience in antibody selection and Jess assay development and optimisation, guarantees reliable target detection, tailored to your unique project requirements.
The key advantages of working on the Jess platform for analysis are:
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Automated protocol with optimised reagent and precedented antibodies provide improved precision, reproducibility and consistent results for in-depth protein characterisation
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Quantitative protein data from low sample volumes – well suited for the availability and diversity of preclinical research sample types e.g. from cell-based assays
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Multiplexing capabilities allow simultaneous target detection and analysis – great for on-board normalisation and evaluation of target pairs e.g. total and phosphorylated targets
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GxP-compliant software for regulatory compliance
Gene expression – quantitative, target specific data
As an early adopter of qRT-PCR back in the 1990s, Pharmagene has continued to support the expression analysis of specific targets in human tissues.
We maintain our high-quality, well annotated database of qualitative “whole body” human gene expression profiles, interrogate expression of disease-specific targets across tissue cohorts and utilise qRT-PCR endpoints in many cellular biology studies. It remains our go-to platform for specificity and sensitivity and over the years we have supported many different types of project – from biodistribution of oligonucleotide/ASO drug candidates to characterisation of gene changes in disease and treatment impact.
Core capabilities
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Custom assay development for specific gene targets and transcripts
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High-quality RNA and DNA extraction and QC from human biosamples (e.g., biofluids, tissue, hair follicles, primary cell models)
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Quantitative gene expression analysis using RT-qPCR
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Sample preparation for other downstream applications (e.g., NGS, biobanking)
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GLP/GCLP compliant laboratory operations
Applications
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Human tissue and biofluid-based target and biomarker validation
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Gene expression changes in response to drug treatment in samples from in vitro and in vivo studies
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Mechanism of action (MoA) studies
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Biodistribution studies (e.g. antisense oligonucleotide therapy, gene therapy, virally delivered therapeutics, miRNA therapy)
XpressWay Profile example: quantitative gene expression data for a G-Protein Coupled Receptor
Data are expressed as target Copy number (Cn) per 100ng of total RNA and are presented as the mean data from 3 individual donors of each of the 72 different human tissues that comprise the profile. The stellar plot is presented on a log scale, with low or no expression in the centre of the plot and increasing expression towards the outer edge of the plot.
Whole body target expression – XpressWay™ Profiles
Understanding how a therapeutic candidate may impact different systems throughout the human body can help to make more informed development decisions.
Using a set of 72 non-diseased human tissues and a validated quantitative reverse transcription-polymerase chain reaction (qRT-PCR) methodology to profile gene expression, Pharmagene has developed XpressWay™ Profiles, a robust platform to provide insight into whole body target expression.
Pharmagene's established XpressWay Profile database contains profiles for more than 2,300 genes, which enables immediate access to robust data for your target(s) of interest. In addition, we can rapidly generate these data profiles for new targets and/or variants on demand.
XpressWay™ Profiles deliver data on how therapeutics may impact tissue and organ systems
Data are generated using 72 non-diseased tissues representing all core organ systems, each tissue from n=3 independent donors.
This helps provide both a broad and deep understanding of potential physiological impact of a candidate therapeutic.
XpressWay Profile example: quantitative gene expression data for a G-Protein Coupled Receptor
Data are expressed as target Copy number (Cn) per 100ng of total RNA and are presented as the mean data from 3 individual donors of each of the 72 different human tissues that comprise the profile. The stellar plot is presented on a log scale, with low or no expression in the centre of the plot and increasing expression towards the outer edge of the plot.
Applications
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Target validation
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Safety liability
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Drug repurposing
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Validation of animal models
Advantages
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Whole body profiles using well characterised, non-diseased human tissues
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qRT-PCR based for sensitive and quantitative expression data
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Validated and robust methodology
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Off the shelf access - existing profiles can be leveraged for rapid data delivery, or new profiles can be readily generated