Challenges in the Detection of Metabolic Biomarkers Using a Multi Plex Assay

In the work we present to the 14th EBF Open Symposium, we show a 5 Plex method for the detection of metabolic biomarkers in human plasma was developed using the U-PLEX technology on the MSD platform.

The complexities surrounding this method were the establishment of endogenous QCs for each chosen biomarker and evaluating the potential use and need for buffer QCs vs matrix QCs where applicable. Additional challenges were faced while trying to ascertain a single minimum required dilution to enable accurate quantification of all biomarkers simultaneously, encompassing the different detection ranges and limits for each biomarker.

The method development focused on the challenges of balancing the detection of the 5 biomarkers; ensuring that each could be quantified within its own required range whilst maintaining a method that was fit for purpose for all 5 biomarkers. To enable this balance, the context of use for each biomarker was evaluated and the method adapted as required.

A fit for purpose method was established that met the requirements for the need and context of use. The method was used successfully in the quantification of human samples from a clinical study.

You may find below the poster for your ready reference:


02/12/2021 10:27

Considerations to properly assess drug stability within biological samples

Assessing drug stability during method development and validation is of paramount importance. The concentration of the target analyte must remain unaffected throughout the lifecycle of the samples to ensure the reliability of the assay data. However, a decrease in analyte concentration in a biological fluid is not always due to a lack of stability.

14/06/2021 15:46

Challenges in development of robust analytical methods for the quantitation of low molecular weight compounds by LC-MS/MS

Method development can be considered one of the most challenging task in a Bioanalytical laboratory. When working with LC-MS/MS and low molecular weight drugs, low degree of ionization, poor fragmentation, higher presence of isobaric compounds or analyte loss due to their high volatility can get in the way of a successful method development.

04/06/2021 11:22

Qualification of a method for the measurement of a potentially therapeutic protein

A recent project completed by Anapharm involved qualifying an analytical method for the measurement of a therapeutic protein in human skin tissue. The protein of interest is one that is expressed ubiquitously in a number of tissues and cell types.