Project focus

Boehringer Ingelheim is a global pharmaceutical company committed to research and development, manufacturing and marketing of novel human and veterinary medicines. A crucial step in the development of new therapies is evaluating the properties of potential drugs.

The Challenges

We analyze plasma, urine, and tissue samples from human and animal studies, looking for in vivo biotransformation of proteins, and quantifying both proteins and oligonucleotides. The biggest challenge for these analyses is sample clean-up and enrichment to maximize sensitivity. For example, we routinely use immunocapture to purify and enrich protein biologics from plasma samples prior to MS/MS quantification on a SCIEX Triple Quad™ system, achieving sensitivity in the region of low nanograms per milliliter.

The Solution

Our department has a dozen SCIEX mass spectrometers. These include the SCIEX Triple Quad systems that we have used for many years, and the more recently acquired high-resolution QTOF systems—three TripleTOF® 6600 LC-MS/MS Systems. These systems are used extensively for preclinical analysis of small and large molecules and oligonucleotides for a wide variety of applications, including immunotherapies, cancer therapies, cardiovascular disease, and metabolic disorders. We rely on the QTOF systems for simultaneous quantification and metabolic profiling of oligonucleotides. The high resolution and mass accuracy of these systems allow us to reduce, or even eliminate, the effect of any interfering peaks and improve sensitivity.

The Outcomes

We have shown that mass spectrometers can now match or even exceed the sensitivity of fluorescence techniques, achieving up to 10 times more sensitivity using MS/MS analysis. Typically, we reach a sensitivity of less than 10 nanograms per milliliter using high-resolution MS quantification—as low as picograms per milliliter using MS/MS—which we think could be better than most liquid chromatography (LC)-fluorescence methods or enzyme-linked immunosorbent assay (ELISA) measurements. At the same time, we have been able to halve the runtime from roughly 10 minutes a sample by LC-fluorescence to approximately 5 minutes by MS, increasing throughput significantly. Even more of an advantage is that the QTOF systems allow us to quantify the parent oligonucleotide and simultaneously perform metabolite profiling and quantification, which is hard to achieve by LC-fluorescence, and not possible with ELISA or quantitative polymerase chain reaction (qPCR) measurements.