Test tubes in a laboratory

In Vitro Sampling

Identify prospective VOC biomarkers relevant to key disease processes through in vitro analysis


Linking breath and biology

Discovering and validating biomarkers for disease is a key stage in developing valuable and effective diagnostic tests for clinical applications. You may already have a detailed understanding of the biology of your disease of interest but how can you go about translating that into the clinic?

If you’ve already conducted a clinical trial, in vitro sampling could be an efficient route to achieving the level of validation you need to achieve regulatory approval and widespread acceptance.

Our in vitro sampling pipeline supported by our Breath Biopsy® Services can help you to identify volatile organic compounds (VOCs) produced by your laboratory samples that have the potential to be effective biomarkers for non-invasive clinical breath testing. We provide an optimized detection process supported by expert VOC analysis and identification that provides high confidence candidate biomarker discovery ideal for clinical translation.

in vitro pipeline

Our approach is designed to bridge the gap in biomarker research. Some existing studies have investigated metabolic processes involved in disease but have not identified clinically viable biomarkers. While others have taken an untargeted approach to discovering clinical biomarkers without insight from the underlying biology.

Many studies show that VOCs on breath have the potential to be valuable biomarkers for a range of diseases. The majority of these have hypothesized biological origins for biomarkers but relatively few have directly investigated the relationship between disease biology and biomarker abundance.

Analyzing VOCs produced by in vitro models of disease or drug response can demonstrate mechanistic links between disease biology and biomarker candidates. These can subsequently be verified in clinical studies sampling the same VOCs on breath, providing vital evidence that could bring diagnostic breath tests into clinical practice.

Why do in vitro sampling?

  • Discovery

    Identify clinically relevant candidate breath biomarkers

  • Disease Relevance

    Directly relate disease biology to breath biomarkers

  • Mechanisms

    Gain insight into mechanisms underlying disease and drug response

  • Cause & effect

    Dissect markers related to causes and effects of disease

  • Validation

    Validate the chemical identities of your biomarkers

  • Regulatory Approval

    Generate evidence to support your case for clinical approval

  • Relevance

    Relate your findings to other published literature

Why is understanding the biological origins of VOC biomarkers important?

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Cells and VOCs
In Vitro flowchart in the Breath Biopsy Laboratory

Breath Biopsy in vitro sampling

Our in vitro Sampling capability integrates seamlessly with Breath Biopsy Services. The Breath Biopsy Laboratory is equipped with in-tube extraction (ITEX) devices for the automated collection of VOCs produced by in vitro cell cultures or macromolecular isolations.

ITEX is coupled to our high-resolution gas chromatography mass spectrometry (GC-MS) biomarker analysis platform. The Breath Biopsy Platform, built around GC-Orbitrap devices, has been optimized for the detection of volatile compounds found on breath and provides high sensitivity VOC detection, even for low-abundance compounds in the parts per trillion (ppt) range.

Key features:

  • VOC collection using automated ITEX dynamic sampling
  • Direct from sample collection to HRAM GC-MS
  • Suitable for study of cell cultures, ex vivo tissues and/or macromolecular isolations
  • Compare disease site VOCs to breath samples on the same platform

Contact us to discuss integrating Breath Biopsy into your research:

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Identifying Translational Biomarkers using Breath Biopsy in vitro Headspace Analysis

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Case Study

Case Study: VOC production from lipid peroxidation

Lipid peroxidation is a biological mechanism associated with many diseases as well as inflammatory immune responses. Cells under oxidative stress produce increased numbers of reactive oxygen species (ROS), which react rapidly with other molecules in the cell including proteins, DNA and lipids.

When ROS react with unsaturated fatty acids from lipids, this triggers a lipid peroxidation reaction that produces a range of VOCs including simple aldehydes and alkanes. The range of VOCs produced means that lipid peroxidation has been suggested as a source of prospective biomarkers in many studies.

We are currently performing in vitro analysis of lipid peroxidation of various polyunsaturated fatty acids to understand the VOCs produced by each lipid species. Using preparations of isolated fatty acids undergoing lipid peroxidation, we’ve shown that each fatty acid produces a different complement of VOCs and that these can be detected and quantified using the Breath Biopsy Platform.

About Lipid Peroxidation

Lipid Peroxidation Case Study
Top – general lipid peroxidation mechanism from fatty acids to VOCs. Bottom – Schematic of superimposed chromatogram peaks for production of a single VOC in a control sample (grey) and after 2 (purple) and 4 (cyan) hours of lipid peroxidation. Charts show differences in VOC production from lipid peroxidation reactions of different fatty acids.
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