Bioanalysis and Immunogenicity Series - Volume 2

04 Sep 2018

Using Innovative and Non-Conventional Techniques to Address Bioanalytical Challenges

The innovative use of bioanalytical technologies is key to overcoming challenges presented by the chemistry and biology of analytes. Non-conventional approaches are part of a diverse range of analytical techniques that are being utilized at Intertek to meet the demands of increasingly complex bioanalytical challenges. Below is a versatile set of techniques to consider.

Liquid Chromatography-High-Resolution Accurate Mass Spectrometry (LC-HRAMS)

LC-HRAMS based protein analyses are increasingly necessary for drug development. The methodologies offer notable strengths for biologics analyses and basic discovery including an increased understanding of biomarkers. LC-HRAMS can be used for product characterization and examination of biomarker panels. It's an option for targeted quantification and higher throughput and it enables intelligent "charting of the course" through the discovery of potential drug targets and biomarkers.

HRAMS is highly reproducible, and among other capabilities, allows analysis of intact protein mass(es). Additionally, and as a complementary technique, peptide mapping is also required for biologic characterization. It provides highly reproducible results and delivers rapid and precise lot comparisons, for example.  It can be used to detect amino acid sequence variants and analyze post-translational modifications.

Nuclear Magnetic Resonance Spectroscopy (NMR)

NMR can support robust methods, compliant with the bioanalytical method validation guidelines for the analysis of a drug's pharmacokinetic properties. Many biologics are PEGylated to increase their half-life resulting in the potential to decrease dosages, which can lead to improved patient experiences with fewer side effects in the body. Regulators are interested in the accumulation of free and drug-bound poly(ethylene glycol) (PEG) in tissue following chronic administration of a drug.

NMR methods have been validated for the detection of PEGylated compounds within the low ug/mL range and across a diverse range of complex biological matrices. Higher-throughput sample analysis can also be supported using instruments with high magnetic field and cryoprobes.

Inductively Coupled Plasma – Mass Spectrometry (ICP-MS)

Metal-containing drugs are currently used to treat cancer, arthritis, diabetes and cardiovascular diseases. ICP-MS can measure metals in preclinical and clinical samples at very low detection limits, in complex biological matrices.

An example of how this technology has be used include measuring Silver contained in medical devices, creams and wound dressings. Validated Bioanalytical methods have been generated for Silver across different matrices including serum, skin pleural fluid and plasma with an LLOQ of 0.2 ng/mL.

The benefits of ICP-MS include exceptional sensitivity; application across diverse sample matrices; compliant methods that are reasonably straightforward to develop and validate; and simultaneous quantification possible for multiple elements in clinical samples. As with any method, there are challenges to consider, such as endogenous levels of metals in biological fluids, also the preparation and analysis process is destructive to the sample.

Selected Ion Flow Tube –Mass Spectrometry (SIFT-MS)

SIFT-MS separates compounds through gas-phase ion-molecule chemistry. Soft chemical ionization helps to overcome challenges typically experienced using conventional chromatography approaches, including challenges with compounds such as formaldehyde, formic acid, and ammonia.  It uses three fundamental components: reagent ion generation and selection (selected ion); analyte ionization (flow tube); and analyte detection (mass spectrometry).

SIFT-MS uses collision rate kinetics to quantitate analytes in real time. It is inherently linear (up to a point) and does not require conventional calibration or standard prep. It offers real-time breath-by-breath analysis, is highly immune to humidity, highly sensitive and offers a wide linear range. Additionally, it offers direct, non-invasive breath analysis without any sample preparation, as well as analysis of diverse compounds in breath, including carbon dioxide.

Bioanalytic testing can present challenges, but taking a novel approach and using pioneering techniques can help. For more insights into innovative, non-conventional testing techniques, download our complimentary webinar recording.