2017 New Investigator Award: Christian Lanshoeft

Christian Lanshoeft (University of Strasbourg / Novartis, Switzerland)

What made you chose a career in bioanalysis?

The initial contact was created during my MSc thesis exploring ultrafast chromatography-free mass spectrometry (MS) based assays for small molecule quantification in clinical samples. Since then, I was fascinated by the diversity associated with the multidisciplinary bioanalytical field. The major driving forces to continue my research in bioanalysis were the broad variety of analytical platforms as well as analytes including their in vivo generated metabolites and I was inspired by the development of new strategies to elucidate biological mechanisms. Consequently, I decided to pursue a PhD in Analytical Chemistry focusing on quantitative MS analysis of (next-generation) biotherapeutics in highly complex matrices.

Describe the main highlights of your bioanalytical work?

Monoclonal antibody (mAb)-related therapeutic proteins including human immunoglobulin G (hIgG), bispecific mAbs or antibody-drug conjugates (ADCs) are the fastest growing therapeutic class within the last decade. As the pharmaceutical industry significantly invest in the development of such constructs, the first part of my industrial PhD thesis, a joint collaboration between Novartis and the University of Strasbourg, focused on the development of generic quantitative MS-based assays at the peptide level using either immunocapture (IC; Lanshoeft et al, Bioanalysis, 2016) or high-resolution MS (HRMS; Lanshoeft et al, Anal Bioanal Chem, 2016) to support pre-clinical trials. Moreover, a high flexibility at the peptide level was demonstrated while exchanging different animal matrices or spiked analytes with each other (Lanshoeft et al, J Pharm Biomed Anal, 2016).

However, the major breakthrough to date was hybridzing IC with HRMS for multiplexed hIgG1 quantification directly at the intact protein level (Lanshoeft et al, Anal Chem, 2017). New levels of bioanalytical approaches can be reached by transferring this approach to next-generation biotherapeutics including ADCs, since qualitative information (drug load distribution or ADC biotransformation) can be provided in parallel with quantitative data (total antibody, total ADC, individual ADC species bearing different amount of payloads) in one combined assay.

How has your work impacted your laboratory, the bioanalytical field and beyond?

The greatest impact lies definitely in the wide applicability of the developed automated generic quantitative MS-based assays. Since the bioanalytical toolbox includes besides IC also direct serum digestion approaches, rapid method development is guaranteed as no tedious/expensive capture antibody development is required. Consequently, quantitative data and structural information (incorporation of several peptides from different mAb regions) can be provided in a timely manner after new mAb-related entities have entered the bioanalytical laboratory.

Additionally, these generic workflows can be applied along the whole discovery/development phase from candidate selection to late stage pharmacokinetic (PK) assessment and can serve as a starting point for any research group or CRO entering the field of quantitative mAb analysis.

Lastly, a high flexibility is also associated with such assays as method transfer between different matrices (serum and tissue) or from pre-clinical to clinical samples is readily possible by generic capture antibody replacement with the mAb target.

Describe the most difficult challenge you have encountered in your scientific career and how you overcame it?

Due to the complexity of lysine conjugated ADCs, low signal intensities were obtained in the full-scan MS spectrum during intact analysis. Thus, a sophisticated step-by-step optimization of the sample preparation resulting in high extraction recoveries with subsequent HRMS optimization was required in order to achieve the demanded sensitivity under normal flow conditions.

Increasing the initial sample volume for extraction was no option as the provided volume from pre-clinical trial should be sufficient for several analytical assays. Neither sample dilution nor adapting the incubation time increased the overall ADC recovery. Thus, a tritium-labeled ADC was used to optimize the IC from serum as other analytical technologies were either not compatible or sensitive enough. In the end by changing the capture antibody, the ADC extraction recovery could be increased providing enough material loaded on column for intact ADC analysis over the whole PK profile.

However, always keep in mind what type of entity should be covered with the bioanalytical assay (e.g. total antibody or total ADC) and what consequences are caused by simply changing a parameter. Thus, a good understanding of the whole sample preparation process for per se complex biotherapeutics at any time is mandatory which could be quite challenging.

Describe your role in bioanalytical communities/groups?

As a third year PhD student, I have already built up a profound and highly interdisciplinary network which does not only include parties from academia and pharmaceutical industry but also diverse MS vendors. These networks were quite often set up during attended international conferences where I consistently present my latest results to a broader scientific audience (e.g. upcoming HPLC2017, ASMS2016, HPLC2015).

Due to this close collaboration and my valuable expertise in the field of protein analysis using (HR)MS, I am often sharing encountered bioanalytical challenges at round-table discussions or numerous user meetings (e.g. Waters’ ADC and Protein Symposium or Thermo’s Biopharmaceutical Characterization Seminar) in order to bring the bioanalysis especially of next-generation biotherapeutics further.

Besides this, I am also giving external lectures to rather unexperienced scientists entering the field of protein analysis by MS e.g. HRMS quantification workshops at my former university (University of Applied Sciences Northwestern Switzerland).

Regarding departmental teaching activities, I was also actively involved in supervising students during their industrial BSc or MSc thesis in terms of assistance during MS analysis. In order to extend my analytical knowledge besides my own research, I am acting on a regular basis as reviewer for various scientific journals (e.g. Anal Bioanal Chem).

List up to five of your publications in the field of bioanalysis:

1 Lanshoeft C, Cianférani S, Heudi O. “Generic hybrid ligand binding assay liquid chromatography high-resolution mass spectrometry-based workflow for multiplexed human immunoglobulin G1 quantification at the intact protein level: application to preclinical pharmacokinetic studies“. Anal Chem, 2017, 89(4), 2628-2635.

2 Lanshoeft C et al. “The flexibility of a generic LC-MS/MS method for the quantitative analysis of therapeutic proteins based on human immunoglobulin G and related constructs in animal studies”. J Pharm Biomed Anal, 2016, 131, 214-222.

3 Lanshoeft C, Heudi O, Cianférani S, Warren AP, Picard F, Kretz O. “Quantitative analysis of hIgG1 in monkey serum by LC-MS/MS using mass spectrometric immunoassay”. Bioanalysis, 2016, 8(10), 1035-1049.

4 Lanshoeft C et al. “Analysis of small molecule antibody-drug conjugate catabolites in rat liver and tumor tissue by liquid extraction surface analysis micro-capillary liquid chromatography tandem mass spectrometry”. Rapid Commun Mass Spectrom, 2016, 30(7), 823-32.

5 Lanshoeft C, Heudi O, Raccuglia M, Leuthold LA, Picard F, Kretz O. “Ultrafast quantitative mass spectrometry based method for ceritinib analysis in human plasma samples from clinical trial”. Bioanalysis, 2015, 7(4), 425-35.

 

Find out more about this year’s New Investigator Award, the judging panel and the rest of our nominees.