BioanalysisAhead of Print Special FeatureFree AccessWRIB Poster Awards winners 2022Chong Kim, Magali Carcenac, Hua-Chen Chang, William McAuliffe & Moo-jin SuhChong Kim*Author for correspondence: E-mail Address: chong.kim@astrazeneca.comAstraZeneca, Gaithersburg, Maryland, United StatesSearch for more papers by this author, Magali CarcenacSanofi, Montpellier, Occitanie, FranceSearch for more papers by this author, Hua-Chen ChangLabcorp Drug Development, Indianapolis, Indiana, United StatesSearch for more papers by this author, William McAuliffeTakeda Development Center Americas, Cambridge, Massachusetts, United StatesSearch for more papers by this author & Moo-jin SuhAstraZeneca, Gaithersburg, Maryland, United StatesSearch for more papers by this authorPublished Online:24 Mar 2023https://doi.org/10.4155/bio-2023-0048AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit Keywords: bioanalysisbiomarkerscell therapygene therapyimmunogenicityvaccineWRIBIntroductionThe 16th edition of the Workshop on Recent Issues in Bioanalysis (16th WRIB) was held on 26–30 September. Over 1000 professionals representing pharma/biotech companies, CROs, and multiple regulatory agencies convened to actively discuss the most current topics of interest in bioanalysis. The 16th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week in order to allow exhaustive and thorough coverage of all major issues in bioanalysis, biomarkers, immunogenicity, gene therapy, cell therapy and vaccines.Moreover, in-depth workshops on ICH M10 BMV final guideline (focused on this guideline training, interpretation, adoption and transition); mass spectrometry innovation (focused on novel technologies, novel modalities, and novel challenges); and flow cytometry bioanalysis (rising of the 3rd most common/important technology in bioanalytical labs) were the special features of the 16th edition.As in previous years, this year's WRIB continued to gather a wide diversity of international, industry opinion leaders and regulatory authority experts working on both small and large molecules as well as gene, cell therapies and vaccines to facilitate sharing and discussions focused on improving quality, increasing regulatory compliance, and achieving scientific excellence on bioanalytical issues.Bioanalysis and Bioanalysis Zone are very proud to continually supporting the WRIB Poster Awards, and we feature the profiles of the authors of the winning posters. Visit www.bioanalysis-zone.com to see the winning posters in full.Development of PK & ADA methods for trispecific antibody: comparison of technologies & optimizationMagali Carcenacmagali.carcenac@sanofi.comPlease can you explain about your role at Sanofi?I am a team leader in the Biological Analysis group within the Biomarkers and Clinical Bioanalysis department at Sanofi, Montpellier (France). Our portfolio presents a large diversity of biological projects that require innovative therapeutic approaches in different indications. Our group develops and validates analytical methods in PK, immunogenicity and biomarker assays in order to assess preclinical and clinical samples in this context.How does it feel to win the 16th WRIB Poster award?I was very proud to win this award and especially grateful to the WRIB team. I was the ambassador for my group and this award is the achievement of the engagement of the whole team which participated in this amazing and thrilling work. We did celebrate this achievement as it should be.Please can you tell us a little about the work on which the Poster was based?The work consisted in developing pharmacokinetic (PK) and anti-drug antibody (ADA) methods for a trispecific antibody, T cell engager, in support of clinical studies.This project was a new challenge due to the expected criteria (sensitivity, selectivity, robustness and target tolerance). We decided to take the risk to evaluate several technologies present in our laboratory, in order to meet the expectations of the project. Three platforms were assessed: SMCxPro, QuickPlex SQ120 and GyrolabxPlore.For Safety reasons and to comply with Health Authorities recommendations, the ADA method has to be highly tolerant to the free drug. Thus, the PEG and Acidification assay (PandA) was selected but must be optimized according to the characteristics of the trispecific antibody.What were the key conclusions from your research?The current results were also based on strong interaction with the clinical teams. Both PK and ADA methods were used to analyze patient samples in the current clinical study. Data recorded confirmed that methods were successfully developed and implemented for patient target population and coped to the analytical challenges to support this clinical study.What are you looking forward to working on over the next year?We look forward to working on therapeutic molecules in rare diseases and addressing complex analytical challenges to improve patients' lives through the miracles of science.Assessment of immunogenicity in SARS-CoV-2 naïve & convalescent subjects following COVID-19 vaccinationHua-Chen Changhua-chen.chang@labcorp.comPlease can you explain about your role at LabCorp?As an Associate Director of Immunology at Biomarker Solution Center in LabCorp Drug Development (a contract research organization), my primary role was to work with clients to provide scientific leadership in translational preclinical/clinical studies, assay qualification and validation, contributing to RFP/RFI documents, and responding to inquiries and providing solutions, which ultimately help clients to navigate the testing laboratories that meet the scope and needs for their studies. My job function was also to leverage emerging technologies and platforms for advancement of biomarkers for drug development. It was a privilege to work with the talented and dedicated colleagues in Covance/LabCorp for 7 years, I recently transitioned to a pharmaceutical company (Loxo@ Lilly) where I will continue the mission to advance drug development for improving human lives.How does it feel to win the 16th WRIB Poster award?I still find it unbelievable that I won the 16th WRIB poster award! It's already a great honor to be accepted for our poster presentation. I must confess that I almost clicked the “phishing” button when I received this award email as I'd never expect to be receiving such an accolade. All the memory of this project came back vividly: from the initial internal grant application to the final legal review of the poster. Across multiple teams globally, everyone worked diligently to ensure that all the assays of this project met the highest standard. The commitment from the LabCorp Science Counsel not only supported the execution of this project, but also set a precedent for more innovative research to come. After realizing that I wasn't dreaming, I immediately forwarded this email to my co-authors and colleagues to spread the joy and glory!Please can you tell us a little about the work on which the Poster was based?Current guidelines from the CDC regarding booster shots for COVID vaccines are largely based on the levels of neutralizing antibody against the spike protein. Despite the indispensable role of T helper cells on the antibody production by B lymphocytes, it's less clear about the level of T cell immunity in determining whether the person is adequately protected from the SARS-CoV-2 viruses. In this poster, we set out to assess the immunogenicity in SARS-CoV-2 naïve and convalescent subjects following COVID vaccination by measuring both humoral and cellular immunity. Enzyme-linked immunospot (ELISpot) assay was utilized for detection of antigen-specific T cells in PBMCs following ex vivo stimulation with different peptide pools (individually or in combination), which encompass the entire spike protein of Wuhan strain of SARS-CoV-2 viruses. In addition to the T cell immunogenicity, serum samples collected from these same donors were analyzed for the levels of neutralizing antibody titers using the PhenoSense® pseudovirus assay. Results allowed us to investigate any correlation between the levels of humoral (neutralizing antibody titers) and cellular (IFNg spot counts) immune responses in the vaccinated subjects who have never been exposed to SARS-CoV-2 viruses (naïve group) and those who have been infected with the viruses (convalescent group).What were the key conclusions from your research?A stronger humoral immune response was found in convalescent subjects after vaccination. However, convalescent subjects with a stronger humoral immune response following vaccination do not always have a robust T-immune response. There is no correlation between the levels of neutralizing antibody and spike-specific T-cell responses in subjects participating in this study. Results provide insights to further understand the spectrum of immunity elicited by COVID vaccines.What are you looking forward to working on over the next year?I'm excited about the precision medicine that promises to advance and revolutionize the treatment of cancer. I'm looking forward to continuing learning the progress of molecular programs (large and small) for targeted cancer therapy. I'd like to explore new tools and approaches to further investigate the drug resistance on a single cell level, and to identify modalities that can overcome such limitations, which could ultimately result in a more efficacious regimen and a better outcome. In addition to research, I'd like to understand the FDA regulations for targeted therapy and companion diagnostics (CDx). Not only for approval of a targeted therapy by the FDA, but also how to utilize the CDx criteria to delineate the underlying biology of tumor burden and cancer types, which can be more informative for effective treatment.Development of a machine learning model for guiding the development of anti-drug antibody assaysChong KimChong.Kim@astrazeneca.comPlease can you explain about your role at AstraZeneca?I serve as an outsourcing specialist and bioanalytical lead at AstraZeneca. I support all phases of drug development – pre-clinical drug discovery and GLP studies, phase 1–3 clinical studies, and regulatory submissions. I specialize in PK and ADA ligand-binding assays.How does it feel to win the 16th WRIB Poster award?It was indeed my greatest honor to win this poster award. This was my first time attending WRIB and I was very impressed by the science and expertise in the bioanalytical community. I am thankful to be part of this wonderful group of scientists and make contributions.Please can you tell us a little about the work on which the Poster was based?Anti-drug antibody assay development is probably one of the most challenging aspects of large molecule bioanalysis. Depending on the assay format and quality of reagents, we often see markedly different assay performance that requires a series of troubleshooting and optimization experiments. In most cases, we rely on our intuition to design the experiments and assay conditions. However, such an approach tends to be time-consuming and inefficient and depends heavily on the experience of the scientist.The goal of our project was to use machine learning to speed up the ADA assay development process. We used a large set of data (∼2500 data points) that we had generated across 34 different method development projects. We initially fed the machine with all the data we had, but it failed to understand what these data points meant. To help the machine understand our data better, we created a new variable called DAC (Detectable ADA Concentration) based on various assay parameters such as master-mix concentrations and MRD. This variable, paired with the Boosted Ensemble of Decision Tree model, showed great potential in predicting ADA assay performance under various conditions.What were the key conclusions from your research?Our model was able to use the input parameters to predict the outcome of each assay condition and predict assay characteristics such as drug tolerance and sensitivity. Such in silico data exploration can help shorten the time-consuming assay ADA assay development.What are you looking forward to working on over the next year?While machine learning is quite popular in the pharmaceutical industry, its use in bioanalysis has been quite limited. In this regard, this project was a great starting point because we saw very promising results using real datasets. The next step is to validate and improve the model. We are working on identifying more sophisticated input parameters (e.g., antibody affinity) to boost the predictive performance of the model. In addition, we are using our new datasets to validate the model. Exciting updates are coming up in the future.Qualification of a Machine Learning Approach for the Analysis of High-Dimension Immune-Profiling Data in Celiac DiseaseWilliam McAuliffeWilliam.McAuliffe@takeda.comPlease can you explain about your role at Takeda?As a scientist at Takeda, I support clinical biomarker strategies in various aspects of drug development and therapeutic areas, specifically through the development of biomarker methods in collaboration with our translational teams. It's an exciting time in drug development with many new modalities and opportunities to make an impact on patient health. And in this field, I can apply my interest in translational work to advancing bioanalytical/biomarker approaches to drug development.How does it feel to win the 16th WRIB Poster award?Honestly, I was surprised at first and now I am very thankful for the opportunity to share this project with the broader bioanalytical community. I suspect there may be many of us in this field challenged with the ever-increasing complexity of datasets derived from our studies. So, I was thrilled to learn this work is of interest to others and can potentially be a useful example more broadly.Please can you tell us a little about the work on which the Poster was based?At times we face challenges with managing the analysis of our high-dimension (i.e., large, and complex multiparameter) datasets derived from our clinical translational efforts.For example, one of the many biomarker platforms we utilize involves a type of single cell cytometry that allows us to identify and resolve the myriad of immune cells that may be present in our clinical trial patient samples more fully. By using this platform, we may gain insight into potential biomarkers of disease state or therapeutic effect.Technically speaking, this platform is an immunophenotyping method called CyTOF which allows us to assess many more cellular identification markers than traditional approaches. But due to the high-dimension nature of the resulting dataset challenges can arise in the analysis of the data.In this poster we give an example of our efforts to assess whether machine learning (ML) can help to explore the full potential of complex datasets such as from CyTOF.We applied ML approaches to CyTOF immune-profiling data from two independent, well-characterized Takeda celiac disease (CeD) clinical studies.By taking this approach of using familiar controlled study data, we were able to ‘qualify’ the ML process to give us more confidence in the results and guide us along the way.What were the key conclusions from your research?This study demonstrated that the FlowSOM machine learning algorithm may be a viable approach to interrogate high-dimension data sets. We were able to qualify this specific ML process and results by confirming known controls in the existing dataset.Machine learning may be used as a complementary approach, secondary to hypothesis driven manual gating, to confirm immune subset findings and to yield deeper cell subset characterization not evident with traditional approaches.What are you looking forward to working on over the next year?I am looking forward to working on progressing some of our programs by potentially applying some of our learnings on ML. For example, We now have an effective ML process example on hand to use for any future high-dimension immune profiling studies. And this is very helpful, because although the poster did not get a chance to illustrate this fully, there was substantial pre-study effort to screen out other non-viable ML approaches that did not work out quite as well for this application, for various reasons.Some interesting immune findings that were derived only from this ML exercise, and not traditional approaches, may be worthy of follow-up in future studies and may help us gain insight into celiac disease.A Novel Hybrid LBA-micro-flow LC–MS/MS assay for PK Bioanalysis of ADCsMoo-Jin Suhmoo-jin.suh@astrazeneca.comMoo-jin Suh is an Associate Director in the Integrated Bioanalysis Group, Clinical Pharmacology and Safety Sciences, at AstraZeneca in Gaithersburg, MA (USA). Moo-jin obtained his PhD degree in Chemistry from the University of Cincinnati (OH, USA) and has 2.5 years of postdoctoral training in pharmacology, at Weill Medical College of Cornell University (NY, USA). Prior to joining AstraZeneca in 2021, Moo-jin built his expertise in proteomics, infectious disease, and antibodies in research and industry sectors (e.g., J Craig Venter Institute, US Army Medical Research Institute of Infectious Disease, and Startup biotech).Moo-jin's expertise lies in high-throughput discovery bioanalysis, mass spectrometry, proteomics, structure elucidation of small molecules, imaging mass spectrometry, characterization of antibodies, and biomarkers in complex biological matrices. Moo-jin has authored more than 28 peer-reviewed publications and book chapters in the areas of Analytical Chemistry, Infectious disease, and Proteomics.Please can you explain about your role at AstraZeneca?As an Associate Director within AstraZeneca's Integrated Bioanalysis group, I am responsible for setting up the bioanalytical strategy and for developing and optimizing assays for the pharmacokinetic, pharmacodynamic (PD), and biotransformation studies of biotherapeutics to support drug program transition through lead selection and into clinical trials. I work closely with my team and other staff within AstraZeneca for PK/PD studies for biotherapeutics (T cell engager, and antibody-drug conjugate) and for exploring potential relationships with safety and efficacy.How does it feel to win the 16th WRIB Poster award?It was an honor to be selected as a recipient of the 16th WRIB poster award. When I heard that I had been awarded, I was quick to inform those who made this possible (Josh Powers, Casey Daniels, Yuling Wu) and we were all thrilled to receive this prestigious award. I feel honored to have our work be selected and can't wait to use the feedback I got in order to better my work in the future. I would like to thank you all for giving me this opportunity, thinking of it as an encouragement to work harder.Please can you tell us a little about the work on which the Poster was based?It has been an outstanding decade for the field of antibody drug conjugates (ADCs), with the approval of more than 8 new ADCs. With the ongoing evolution of ADCs portfolio utilizing site-specific conjugation and a small amount of highly cytotoxic payload, and administering a lower dose of ADCs, more sensitive bioanalytical platforms are highly desirable for quantifying major analytes of ADCs to comprehensively understand PK behavior of ADCs in vivo. The goal of this study was to demonstrate the feasibility of a hybrid ligand-binding assay/microflow liquid chromatography-tandem mass spectrometry (LBA/μLC–MS/MS) for use in the PK study when sample quantities are precious. After optimization, the total antibody PK data from a hybrid LBA/μLC–MS/MS was compared with that of ELISA acquired from the same samples, demonstrating feasibility of a hybrid LBA/μLC–MS/MS. Furthermore, we demonstrated multiple and diverse analytical assays (total antibody, intact antibody and total ADC) with the hybrid LBA/μLC–MS/MS in the challenges of ADC bioanalysis.What were the key conclusions from your research?Sample preparation is one of the most important steps to achieve good results in bioanalysis. For sensitive instruments such as μLC–MS/MS, appropriate sample preparation methods (Immunoaffinity capture, efficient digestion, solid phase extraction, etc.) suitable for μLC–MS/MS were selected for downstream analysis. The use of a highly sensitive hybrid LBA/μLC–MS/MS in the current study was reported for quantification of total antibody, intact antibody and total ADC in sub-microliter sample volume. Together with ELISA approach, this method provides another powerful and sensitive analytical platform for understanding the fate of ADCs in vivo.What are you looking forward to working on over the next year?It is predicted that the use of a hybrid LBA/μLC–MS/MS will gradually increase in order to obtain comprehensive PK information in biotherapeutic dosed samples. In particular, measuring trace amounts of free payload present in circulation provides important information for toxicity assessment and safety concerns. Furthermore, it is expected that the hybrid LBA/μLC–MS/MS will be readily adopted for experiments requiring high sensitivity (e.g., tumor tissue with limited sample size) and limited samples. The hybrid LBA/μLC–MS/MS can reduce reagent consumption, minimize the use of sample volume, and be suitable for capillary micro-sampling to collect small amounts of biological samples. Therefore, it is anticipated that it may be possible to implement the hybrid LBA/μLC–MS/MS in preclinical PK studies to reduce overall animal use and improve animal welfare.DisclaimerThe opinions expressed in this interview are those of the interviewee and do not necessarily reflect the views of Newlands Press Ltd.Financial & competing interests disclosureThe interviewees have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.FiguresReferencesRelatedDetails Ahead of Print Follow us on social media for the latest updates Metrics History Received 15 March 2023 Accepted 15 March 2023 Published online 24 March 2023 Information© 2023 Newlands PressKeywordsbioanalysisbiomarkerscell therapygene therapyimmunogenicityvaccineWRIBDisclaimerThe opinions expressed in this interview are those of the interviewee and do not necessarily reflect the views of Newlands Press Ltd.Financial & competing interests disclosureThe interviewees have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.PDF download