To help provide insight into the recent article published in Bioanalysis: Bioanalytical workflow for novel scaffold protein–drug conjugates: quantitation of total Centyrin protein, conjugated Centyrin and free payload for Centyrin–drug conjugate in plasma and tissue samples using liquid chromatography–tandem mass spectrometry, we spoke with author Wenying Jian, Senior Principle Scientist at Johnson and Johnson (NY, USA). Wenying explains why she felt this was an important area for bioanalysis and worthy of publication. With 17 years of experience in bioanalysis, Wenying’s research has been focused on protein bioanalysis using LC–MS for approximately 6 years.
“I am currently a Senior Principal Scientist within the department of drug metabolism and pharmacokinetics of Janssen research and development, Johnson and Johnson (NY, USA). I am leading a group to provide bioanalytical support, including small and large molecule therapeutics, biomarkers and other novel scaffolds using LC–MS. I received my Bachelor’s in Pharmacy from Beijing Medical University (China) in 1998, Master’s in Microbiology from the Chinese Academy of Sciences (Beijing, China) in 2001, and a PhD in Pharmacology from the University of Pennsylvania (PA, USA) under the supervision of Dr Ian Blair in 2005. After graduation, I worked for Bristol-Myers Squibb (NY, USA) and then Johnson and Johnson to support various bioanalytical activities in discovery, pre-clinical and clinical studies. My research interest is the application of advanced LC–MS methodologies in detection, identification and quantitation of drugs, metabolites and biomarkers. I have published over 40 peer-reviewed journal papers and book chapters, and co-edited two books: ‘Targeted biomarker quantitation by LC–MS’ Wiley (2017) with Dr Naidong Weng, and ‘Sample preparation in LC–MS bioanalysis’ Wiley (in production) with Dr Wenkui Li and Yunlin Fu. I currently serve on the editorial board of the Journal of Pharmacological and Toxicological Methods and the board of Eastern Analytical Symposium (EAS). I also contribute to the bioanalysis community by giving presentations, lectures and short courses.”
1. What inspired you to work in this field of bioanalysis?
Bioanalysis provides very powerful tools to investigate fundamental biological and pharmacological questions as well as to understand properties and effects of drug candidates for pharmaceutical research and development. For my graduate study in Dr Blair’s lab, I conducted basic research on oxidative stress and its impact on biological systems. Using advanced bioanalytical tools, we were able to identify and quantify oxidative stress-induced modifications on endogenous molecules such as DNA and lipids, which has important implications in human diseases. It fascinated me that we were able to quantitatively analyze in vivo processes with such selectivity and sensitivity. In my industrial career, I was further exposed to all stages of drug discovery and development, for which bioanalysis plays an essential role from beginning to the end. It provides fundamental information regarding the ADME properties of the drug candidates as well their effects on the body. As more and more novel molecules and platforms come through our pipelines, we are presented with ever-evolving challenges of developing innovative approaches of bioanalysis, which is inspirational.
2. What impact would you like to see/expect to see as a result of your publication?
Antibody–drug conjugates (ADC) are a well-established platform, having undergone an abundance of research. However, antibodies as carrier platforms have their own limitations such as heterogeneity, instability and production limitation due to their complex structures. Alternative platform scaffold proteins, such as Centyrin presented in this publication, have smaller molecule size, simpler structure, much improved physiochemistry property and can be produced easily by bacterial expression or chemical synthesis. Scaffold protein–drug conjugates (PDC) present one of the future trends of targeted delivery of therapeutics. In this publication, we presented a simple and specific workflow for quantifying PDCs based on novel scaffold proteins. LC–MS/MS methodology was utilized to quantify total protein, conjugates and free payload to understand linker stability and tissue distribution. I would like to see the same workflow to be adapted for future similar scaffold–drug conjugates and site-specific ADCs.
3. What are the next steps for your research and this field of bioanalysis?
As with ADCs, it is important to understand the ADME properties of PDCs by conducting similar research as that previously done for ADCs. These include, but are not limited to, biotransformation of payload, catabolism, drug-antibody (or protein for PDC) ratio determination, distribution in tumors and relationship between linker stability and efficacy. From a bioanalytical perspective, I would explore intact LC–MS analysis on PDCs, which can provide a more holistic view of the overall fate of PDCs in the body. My research interest in recent years has been focused on intact bioanalysis of proteins for quantitation and catabolite identification. Intact analysis is an important complementary approach for protein analysis to the more commonly utilized bottom-up methodology. With improvement in sample preparation, instrumentation and software, I believe intact protein bioanalysis will play an important role in understanding ADME properties of biotherapeutics.
4. Are there any researchers/projects/technologies that you are watching at the moment, and any you think we should be keeping an eye on?
There are a few areas of research/technologies in the field of bioanalysis that are of particular interest to me: (1) ADME of biotherapeutics – for small molecule drugs, ADME is a relative mature field with well-established in vitro and in vivo models. In comparison, there are still challenges and gaps in the understanding of ADME for large molecules such as the lack of in vitro or in vivo models and limitation on analytical technologies. (2) Target engagement – tissue bioanalysis or imaging study of proteins for understanding distribution of biotherapeutics at target sites. Measurement of free versus total target level to provide important information of target engagement and PK/PD correlation. (3) Biomarkers – challenges still remain for ultrasensitive and specific quantitation of endogenous molecules as biomarkers for drug discovery and development research. (4) Capillary electrophoresis –provides superior separation power but currently has limited application in bioanalysis. I would like to see technique advancements in adapting capillary electrophoresis to biological samples.
5. Do you have any advice for anyone who may be interested in working in this field?
As bioanalysts, our mission is to help biologists and chemists to gain insight into the biological systems and their interaction with endogenous or exogenous molecules. It is important for us to not only master the skills of analysis, but also understand the context of the questions to be answered. It is helpful to equip ourselves with knowledge in biology, biochemistry, pharmacology, as well as drug discovery and development. This is an ever-evolving field and we should always be open to embrace new knowledge, technologies and experience.
Reference: Shi C, Goldberg S, Lin T et al. Bioanalytical workflow for novel scaffold protein–drug conjugates: quantitation of total Centyrin protein, conjugated Centyrin and free payload for Centyrin–drug conjugate in plasma and tissue samples using liquid chromatography–tandem mass spectrometry. Bioanalysis. 10(20): 1651–1665 (2018)