Conference Report | News & Analysis
New setting for the Land O’Lakes bioanalytical conference Fourteenth Annual Land O’Lakes Bioanalytical Conference University of Wisconsin, Madison, WI, USA, 15–18 July 2013 The University of Wisconsin bioanalytical conference is presented each year by the Extension Services in Pharmacy, the professional development department within the School of Pharmacy. The purpose of this 3-day conference was to provide an educational forum to discuss issues and applications associated with the analysis of xenobiotics, metabolites, biologics and biomarkers in biological matrices. The conference was designed to include and encourage an open exchange of scientific and methodological applications for bioanalysis. To increase the interactive nature of the conference the program was composed of a mixture of lectures, interactive discussions, poster sessions and roundtables. This paper summarizes the presentations at the Fourteenth Annual Conference, offered in a new venue.
The Fourteenth Annual Land O’Lakes Bioanalytical Conference, titled ‘Emerging Challenges and Evolving Technologies for Bioanalysis’, was held on 15–18 July 2013. This year’s program offered several changes from previous conferences: moving the meeting site to Madison and improved conference facilities on the University of Wisconsin Campus; shortening of the program by 1 day and adding plenary sessions in the afternoons; and presenting conference material in electronic rather than print format. The program provided an educational forum with presentations by scientific leaders from academia, industry, contract laboratories and regulatory authorities. The conference traditionally brings together an international group of bioanalytical scientists with the express intent of discussing cutting-edge science for small- and large-molecule bioanalysis in a relaxed atmosphere that also promotes networking opportunities. The new venue maintained this informal environment for learning and networking. The success of the program has been due in large part to the work of the Planning Committee, which consists of scientists involved with bioanalysis on a daily basis, and their ability to identify timely topics and presenters who are experts in the field. Specific objectives for the conference, a list of the Planning Committee members and summary of the conference evaluations can be found at [1]. On Monday evening the conference opened with a presentation intended to be scientifically thought-provoking by dealing with a topic not part of the daily bioanalytical environment (also new for 2013). This presentation focused on the role of stem cell technology in drug development
and was presented by Brad Swanson (Cellular Dynamics, WI, USA). Following a brief background on human pluripotent stem cells, he discussed modeling human biology ‘in a dish’ and applications in drug discovery and toxicology, as well as disease biology modeling. He presented challenges and caveats when considering using pluripotent stem cell-derived tissue cells.
10.4155/BIO.13.232 © 2011 Future Science Ltd
Bioanalysis (2013) 5(21), 2601–2605
Three morning plenary sessions Morning meetings consisted of plenary sessions focusing on specific themes associated with bioanalytical issues. The first plenary session focused on the technology and applications for biomarkers. Chad Ray (ICON Development Solutions, NJ, USA) started this session by giving an overview of biomarkers. He presented the rational for measuring biomarkers, provided a brief summary of the roles of biomarkers (diagnostic, prognostic and predictive), and discussed in detail how the biomarkers are applied at different stages of drug development. Ray listed the challenges of developing a precise and accurate biomarker assay, emphasizing the importance of examining the agreement of biomarker results using different methods. He presented examples of biomarker application in drug development. Michael Burczynski, (BristolMyers Squibb, NJ, USA) presented biomarker technologies and analytical method validation in early clinical drug development. The current biomarker technologies include flow cytometry, histochemistry, transcriptomics, and genetics. These technologies enable biomarker-based decision making and also lay the foundation for companion diagnostic development. Burczynski discussed the process of performing analytical
James E DeMuth*1, Stacy Ho2 , Chad Briscoe3, Matthew Cyronak4, Eric N Fluhler5, Qin C Ji6 & Priya Sriraman7 University of Wisconsin, 777 Highland Ave., Madison, WI 53705, USA 2 Sanofi, MA, USA 3 PRA, KS, USA 4 Alliance Pharma, Inc., Malvern, UK 5 Pfizer Research, NY, USA 6 Bristol-Myers Squibb Company, NJ, USA 7 Celgene Corporation, NJ, USA *Author for correspondence: E-mail: jedemuth@ pharmacy.wisc.edu 1
ISSN 1757-6180
2601
News & Analysis | Conference Report validation of biomarker assays, which starts with identifying applicable guidance, followed by asking fundamental questions of the assay (analyte to be measured, sensitivity, precision and accuracy, and so on). He presented a decision flow chart for identifying fit-for-purpose validation requirements, which incorporates the considerations of regional requirements, timeline, reagent availability and resource. He also speculated the future trends in biomarker research. Soma Ray (Biogen Idec, MA, USA) discussed the why, how and practical considerations for evaluating different assay parameters in the context of fit-for-purpose biomarker analysis. She included best and worst cases to facilitate the discussion of standard curves, QCs, parallelism, selectivity, sensitivity and specificity for biomarker assays. She presented the value of using endogenous QCs during the biomarker analysis, which are typically prepared by pooling study samples. She also elaborated on the difference between dilution linearity and parallelism. The session concluded with regulatory consideration by Suresh B Naraharisetti (US FDA, MD, USA). Biomarkers are increasingly used to assess the safety and/ or effectiveness of new drugs and biological products. As a result of this important role, the reliability of the data generated by assays used to measure biomarker concentrations needs to be assured. For the extent of biomarker method validation; a fit-for-purpose approach should be used. For instances in which the biomarker data will support a regulatory action such as determination of safety and/or effectiveness, or support labeled dosing instructions, the assay should be fully validated. In other cases, where the sponsor uses the biomarker data internally, for example, to make decisions about candidate selection or continuing product development, the sponsor can use extent of analytical method validation as it deems appropriate for these decisions. The Wednesday morning plenary session focused on evolving technologies and methods. The session covered application of HRMS to both small molecule and protein therapeutics, design considerations for developing ligandbinding assays (LBAs) for antibody–drug conjugate (ADC) programs, as well as establishing automation in the bioanalytical laboratory. Cynthia Chavez-Eng (Merck, PA, USA) presented a comparison between the triple quadrupole system and HRMS using two case studies. HRMS allowed improved mass resolution and the use of a generic method resulting in reduced development times and post-acquisition data mining. Precision and accuracy were comparable to SRM 2602
Bioanalysis (2013) 5(21)
and were shown to be dependent on using the optimal mass extraction windows. Chavez-Eng discussed the concept of ion-throttling to reduce the number of ions reaching the detector and concluded the benefits of HRMS can be fully realized as the user base expands and software becomes more adaptable in a regulated environment. Yves Le Blanc (AB SCIEX, Canada) described the use of HRMS for the quantitation of peptides and whole proteins. Generating narrow extracted ion chromatograms on major isotopes as well as major charge states, and combining (additive) the information generated were discussed as effective approaches to improved sensitivity. For intact proteins, some sample pretreatment (depletion/enrichment) strategy was required to eliminate major matrix proteins. The high duty cycle of HRMS allows for detection of multiple tryptic peptides, which then allows post-acquisition identification of the most sensitive and selective fragment. Lindsay King (Pfizer, CT, USA) presented design considerations for LBAs that are used in early stages of a program to better understand PK, PD and immunogenicity of drug candidates. Emphasis was on assays to support ADC programs. Two different LBAs to measure ADCs were described. The advantages and disadvantages of each assay were compared as well as the utility of data obtained from each assay. The need to understand the influence of soluble or shed antigen on an LBA was discussed. If target concentrations are dynamic, the utility of an assay that measures both target-bound (total) and unbound (free) drug was highlighted. Incorporation of immunogenicity assessments was useful to better understand anomalous PK even during early stages of a program. Vimal Patel (Amgen, CA, USA) presented operational advantages and challenges of implementing automation in a bioanalytical laboratory. Multiple automated liquid handlers, their ease of use and programming were discussed. Generic scripts, once established, allow improved efficiency, data quality, pass-rate and reproducibility with minimum user input. A case study was presented showing integration of an automated microfluidics platform, a pipetting wizard and a laboratory information management system. A modern off-the-shelf pipetting wizard with a wide range of pipetting protocols and a laboratory information management system interface was adapted to fit sample pipetting and dilutions. Integration of modular applications of existing protocols and interfaces allowed for easy implementation with minimal resources. future science group
Conference Report | News & Analysis The Thursday morning plenary session on the final day featured four talks focusing on timely topics in bioanalysis. Rebbeca Sendak (Genzyme, MA, USA) discussed biosimilars and the challenges presented in order to analytically characterize complex biomolecules and demonstrate biosimilarity from a chemistry, manufacturing and control perspective. Through several examples she showed there is a spectrum of complexity with biomolecules making a single analytical approach impossible. The more complex the biomolecule the less one’s ability to fully perform analytical characterization. Analytical characterization of biosimilars requires a significant amount of planning and investment. Sendak discussed the bioanalytical challenges that biosimilars pose not only for PK analysis but also with evaluating immunogenicity. Surinder Kaur (Genentech, CA, USA) presented a very detailed overview of the analytical considerations for ADCs. ADCs have complex structures that require several methodologies and strategies in order to characterize the PK of the different components. She emphasized that success is highly dependent on extensive collaboration between the small- and large-molecule groups through the integration of ligand binding, LC–MS/MS and hybrid assay strategies and examples were presented. ADCs can also elicit an immune response that presents an analytical challenge because any of the component – antibody, linker and cytotoxic drug – may result in the generation of the antitherapeutic antibody response. Surinder emphasized that it is important to understand ADC structures in vivo to ensure appropriate assays are developed to understand PK and immunogenicity of these drug candidates. Jim Vrbanac (MPI Research, MI, USA) discussed the metabolites in safety testing guidelines as they apply to preclinical and clinical studies and described how high-resolution isotope-dilution MS (HRIDMS) can be a powerful tool in understanding the metabolism of drug candidates early in development. He discussed the use of isotopedilution MS as a precise and accurate technique and the use of HRMS versus MS/MS. Vrbanac presented several HRID-MS test experiments that demonstrated the utility of the technique. Due to recent advances in cell and tissue technologies the use of HRID-MS will continue to evolve. Peter van Amsterdam (Abbott, the Netherlands) presented a thorough comparison of the many regulatory guidelines focusing on bioanalysis. Started with a historical perspective he went on to show where the guidelines future science group
from the European Medicines Agency, FDA, ANVISA (Brazil), and MHLW (Japan) are harmonized and where they differ. His conclusion was that a bioanalytical group performing regulated analysis can begin with the European Medicines Agency guidelines as a base and make modifications from there. van Amsterdam gave a very extensive reference section at the end of his talk that would be useful to anyone delving into regulated bioanalysis. New afternoon sessions Because of the change in venue and shorter schedule, traditional afternoon workshops were replaced with additional plenary sessions. Tuesday afternoon’s program focused on outsourcing issues in bioanalysis from different segments of the pharmaceutical industry. Kenneth Ruterbories (Lilly, IN, USA) provided an overview of his company’s evolution from a fully integrated pharmaceutical company (everything in-house) to a fully integrated pharmaceutical network (a blend of in-house and outsourced analysis). Bioanalysis was identified as essential but not a core function and has evolved be a fully outsourced function. This evolution over 15 years resulted in reductions in instruments and personnel. Ruterbories reviewed the approach to selecting and narrowing down vendors to a small preferred list. The expectations for providers have evolved and these expectations have increased. He hopes to see an evolution in the way methods are developed; using good science but keep the methods simple. He expects CROs to be at the forefront of technology in areas such as eDocumentation and accurate quantification. He concluded that “outsourcing is an investment.” Brian O’Dwyer’s (ICON Development Solutions, NJ, USA) presented the CRO perspective and highlighted the challenges and evolving technologies. He focused on different aspects of managing partnerships with pharmaceutical companies and talked about the various challenges a CRO faces; it is a balance between meeting client’s expectations for a very high level of technical service and very slim operating margins. CROs need to be at the forefront of technology and new techniques while still maintaining profitability. O’Dwyer reported profitability data comparing eight Pharma companies and four CROs demonstrating that there were margins of 22 and 4%, respectively. However, the upside includes a large and growing percentage of bioanalysis work being outsourced. Closer partnerships between CROs and pharmaceutical companies can provide improved service and www.future-science.com
2603
News & Analysis | Conference Report cost savings to the benefit of both parties. Wei Zhou (Novartis, MA, USA) reviewed the challenges of outsourcing offshore. She gave an overview of her company’s philosophy of outsourcing. Novartis has also consolidated their bioanalytical units, but still keeps most work in-house through early-stage development. At that point they use a small number of vendors that have been carefully selected. For GLP studies they keep most studies in North America and Europe due to regulatory and logistical advantages. The cost at these sites is greater, but has decreased in recent years. CROs in China and India are used primarily for cost savings. The performance tends to be similar to other CROs around the world; however, the prices are rising and there are other disadvantages. Challenges in China include language, export of samples and time zones difference; with India there were recent changes in their import laws; and for Jordan and Russia it is primary a lack of familiarity. The Wednesday afternoon plenary session focused on prediction and prevention of immunogenicity. Crystal Sung (Sanofi, MA, USA) discussed the concept of immune tolerance induction (ITI) as it applies to drug development and patient treatment. She described the potential consequences of antidrug antibody formation, including its impact on PK, efficacy and safety. The concept and application of ITI was then discussed in the context of the enzyme replacement therapy used to treat Pompe Disease, a rare and often fatal disease caused by a deficiency of lysosomal acid a-glucosidase. While treatment for Pompe Disease exists, effectiveness is impacted by a high incidence of antidrug antibody formation in certain patient genotypes (e.g., crossreactive immunologic material – in infants). Various approaches to ITI were reviewed and clinical data were presented demonstrating how both prophylactic and therapeutic regimens with immune modulators can be used to obtain long-lived tolerance in Pompe Disease infants. Sung stressed that investigation into the mechanism of these responses is warranted so that translation to treatments for other life-threatening diseases can be understood. Bonita Rup (Pfizer, MA, USA) followed with a discussion on the state of the science for immunogenicity prediction, discussing some of the factors that influence immune responses to biotherapeutics proteins. These included product-related factors (e.g., sequence, aggregation, and post-translational modifications), patient influence (e.g., disease state, genetic factors and immune status) and treatment factor (e.g., regimen, route and co-medications). Rup discussed 2604
Bioanalysis (2013) 5(21)
the array of tools currently available for predicting immunogenicity prior to first-in-human trials, including: in silico T-cell epitope analysis, in vitro HLA binding assays, ex vivo dendritic cell T-cell activation assays, the artificial lymph node, and the concept of a ‘humanized mouse’ model. She described the advantages, limitations and future prospects of each of these tools with an emphasis on applicability. She concluded by emphasizing that many of these tools are in their infancy and require standardization and additional learning. The second part of Wednesday afternoon was the analytical investigators forum, which provided a platform for scientists to present research building on the major themes of the conference. Joy Hoesing (PRA, KS, USA) presented a case study investigation of whole blood sample collection and handling stability. Industry guidance indicates the necessity for evaluating sample collection and handling stability; however, there is no specific guidance on how the stability assessment should be conducted. Two specific whole blood sample collection and handling examples were covered in the presentation. The first case study demonstrated the possible impact of anticoagulant selection on analyte conversion, both in fortified whole blood samples and incurred samples. The second case study investigated whole blood stability in the presence of additional metabolites with results indicating the importance of conversion considerations in relation to both incubation time and temperature. Jonathan Haulenbeek (Bristol-Myers Squibb, NJ, USA) gave an overview of the importance of characterizing critical reagents for LBAs. Detailed attention was paid to defining the molar incorporation ratio of labeled critical reagents. A general process for labeling and subsequent analysis was given. Mass spectral techniques for incorporation ratio were the focus of development and late-phase case studies. The cases studies showed that analysis by MALDI-TOF-MS had aided assay optimization and trouble shooting. Detailed characterization of labeled reagents was shown to make method development and method transfer more efficient. The presentations initiated a lot of interest and discussions among the conference attendees. Opportunities for interacting & networking On Tuesday afternoon, following the plenary session, there were informal roundtables held on a variety of topics. These discussions and facilitators covered: future science group
Conference Report | News & Analysis Best practices of discovery bioanalysis: balancing data quality and productivity (Stacy Ho, Sanofi, MA, USA);
resulted in more interaction and greater camaraderie between participants than in previous years at the resort.
Personal skills development: managing through change in the laboratory (Randall H Guthrie, RHG & Associates, LLC, WI, USA);
Conclusion Due to the informal, noncommercial and improved conference setting, attendees have more time to meet each other, discuss their organizations, practices and the science than they would experience at large international conventions. Feedback was very positive and showed a significant improvement in evaluation questions related to the conference facility, food service and guest rooms. Next year’s conference, which is currently being developed, will follow a similar format at the same conference facility and will be posted on the University’s website when finished [2].
n
n
Dealing with daily bioanalytical laboratory problems (Chad Briscoe, PRA, KS, USA);
n
DBS and microsampling: what does the future hold? (Douglas J Turk, Lambda Therapeutic Research Inc., Canada);
n
Compliance challenges for the bioanalytical scientist (R John Stubbs, Stubbs and HenselPharma Consulting, LLC, PA, USA).
n
A major challenge with the new venue After 54 years of presenting Land O’Lakes Conferences at remote resorts, the major challenge facing the conference organizers was to create an interactive environment and keep people together with the distractions of an urban setting. This was accomplished by adding a new dinner presentation on the opening evening, better break service during the plenary sessions, roundtable discussions in a beer garden at a local brewery, and inclusion of gift cards for participants randomly selected at various times during the poster session. Initial impressions and conference evaluations support that these changes
Disclosure The views expressed in Naraharisetti’s presentation are those of the presenter and do not reflect official policy of the US FDA. No official endorsement by the FDA is intended or should be inferred.
Financial & competing interests disclosure The authors 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.
Websites 1
14th Annual Land O’Lakes Bioanalytical Conference – Emerging Challenges and Evolving Technologies for Bioanalysis. http://ce.pharmacy.wisc.edu/2013JulyLOLInfo
future science group
2
15th Annual Land O’Lakes Bioanalytical Conference (Madison, WI, USA) 21–24 July 2014 . http://ce.pharmacy.wisc.edu/2014JulyLOLInfo
www.future-science.com
2605
New setting for the Land O’Lakes bioanalytical conference Fourteenth Annual Land O’Lakes Bioanalytical Conference University of Wisconsin, Madison, WI, USA, 15–18 July 2013 The University of Wisconsin bioanalytical conference is presented each year by the Extension Services in Pharmacy, the professional development department within the School of Pharmacy. The purpose of this 3-day conference was to provide an educational forum to discuss issues and applications associated with the analysis of xenobiotics, metabolites, biologics and biomarkers in biological matrices. The conference was designed to include and encourage an open exchange of scientific and methodological applications for bioanalysis. To increase the interactive nature of the conference the program was composed of a mixture of lectures, interactive discussions, poster sessions and roundtables. This paper summarizes the presentations at the Fourteenth Annual Conference, offered in a new venue.
The Fourteenth Annual Land O’Lakes Bioanalytical Conference, titled ‘Emerging Challenges and Evolving Technologies for Bioanalysis’, was held on 15–18 July 2013. This year’s program offered several changes from previous conferences: moving the meeting site to Madison and improved conference facilities on the University of Wisconsin Campus; shortening of the program by 1 day and adding plenary sessions in the afternoons; and presenting conference material in electronic rather than print format. The program provided an educational forum with presentations by scientific leaders from academia, industry, contract laboratories and regulatory authorities. The conference traditionally brings together an international group of bioanalytical scientists with the express intent of discussing cutting-edge science for small- and large-molecule bioanalysis in a relaxed atmosphere that also promotes networking opportunities. The new venue maintained this informal environment for learning and networking. The success of the program has been due in large part to the work of the Planning Committee, which consists of scientists involved with bioanalysis on a daily basis, and their ability to identify timely topics and presenters who are experts in the field. Specific objectives for the conference, a list of the Planning Committee members and summary of the conference evaluations can be found at [1]. On Monday evening the conference opened with a presentation intended to be scientifically thought-provoking by dealing with a topic not part of the daily bioanalytical environment (also new for 2013). This presentation focused on the role of stem cell technology in drug development
and was presented by Brad Swanson (Cellular Dynamics, WI, USA). Following a brief background on human pluripotent stem cells, he discussed modeling human biology ‘in a dish’ and applications in drug discovery and toxicology, as well as disease biology modeling. He presented challenges and caveats when considering using pluripotent stem cell-derived tissue cells.
10.4155/BIO.13.232 © 2011 Future Science Ltd
Bioanalysis (2013) 5(21), 2601–2605
Three morning plenary sessions Morning meetings consisted of plenary sessions focusing on specific themes associated with bioanalytical issues. The first plenary session focused on the technology and applications for biomarkers. Chad Ray (ICON Development Solutions, NJ, USA) started this session by giving an overview of biomarkers. He presented the rational for measuring biomarkers, provided a brief summary of the roles of biomarkers (diagnostic, prognostic and predictive), and discussed in detail how the biomarkers are applied at different stages of drug development. Ray listed the challenges of developing a precise and accurate biomarker assay, emphasizing the importance of examining the agreement of biomarker results using different methods. He presented examples of biomarker application in drug development. Michael Burczynski, (BristolMyers Squibb, NJ, USA) presented biomarker technologies and analytical method validation in early clinical drug development. The current biomarker technologies include flow cytometry, histochemistry, transcriptomics, and genetics. These technologies enable biomarker-based decision making and also lay the foundation for companion diagnostic development. Burczynski discussed the process of performing analytical
James E DeMuth*1, Stacy Ho2 , Chad Briscoe3, Matthew Cyronak4, Eric N Fluhler5, Qin C Ji6 & Priya Sriraman7 University of Wisconsin, 777 Highland Ave., Madison, WI 53705, USA 2 Sanofi, MA, USA 3 PRA, KS, USA 4 Alliance Pharma, Inc., Malvern, UK 5 Pfizer Research, NY, USA 6 Bristol-Myers Squibb Company, NJ, USA 7 Celgene Corporation, NJ, USA *Author for correspondence: E-mail: jedemuth@ pharmacy.wisc.edu 1
ISSN 1757-6180
2601
News & Analysis | Conference Report validation of biomarker assays, which starts with identifying applicable guidance, followed by asking fundamental questions of the assay (analyte to be measured, sensitivity, precision and accuracy, and so on). He presented a decision flow chart for identifying fit-for-purpose validation requirements, which incorporates the considerations of regional requirements, timeline, reagent availability and resource. He also speculated the future trends in biomarker research. Soma Ray (Biogen Idec, MA, USA) discussed the why, how and practical considerations for evaluating different assay parameters in the context of fit-for-purpose biomarker analysis. She included best and worst cases to facilitate the discussion of standard curves, QCs, parallelism, selectivity, sensitivity and specificity for biomarker assays. She presented the value of using endogenous QCs during the biomarker analysis, which are typically prepared by pooling study samples. She also elaborated on the difference between dilution linearity and parallelism. The session concluded with regulatory consideration by Suresh B Naraharisetti (US FDA, MD, USA). Biomarkers are increasingly used to assess the safety and/ or effectiveness of new drugs and biological products. As a result of this important role, the reliability of the data generated by assays used to measure biomarker concentrations needs to be assured. For the extent of biomarker method validation; a fit-for-purpose approach should be used. For instances in which the biomarker data will support a regulatory action such as determination of safety and/or effectiveness, or support labeled dosing instructions, the assay should be fully validated. In other cases, where the sponsor uses the biomarker data internally, for example, to make decisions about candidate selection or continuing product development, the sponsor can use extent of analytical method validation as it deems appropriate for these decisions. The Wednesday morning plenary session focused on evolving technologies and methods. The session covered application of HRMS to both small molecule and protein therapeutics, design considerations for developing ligandbinding assays (LBAs) for antibody–drug conjugate (ADC) programs, as well as establishing automation in the bioanalytical laboratory. Cynthia Chavez-Eng (Merck, PA, USA) presented a comparison between the triple quadrupole system and HRMS using two case studies. HRMS allowed improved mass resolution and the use of a generic method resulting in reduced development times and post-acquisition data mining. Precision and accuracy were comparable to SRM 2602
Bioanalysis (2013) 5(21)
and were shown to be dependent on using the optimal mass extraction windows. Chavez-Eng discussed the concept of ion-throttling to reduce the number of ions reaching the detector and concluded the benefits of HRMS can be fully realized as the user base expands and software becomes more adaptable in a regulated environment. Yves Le Blanc (AB SCIEX, Canada) described the use of HRMS for the quantitation of peptides and whole proteins. Generating narrow extracted ion chromatograms on major isotopes as well as major charge states, and combining (additive) the information generated were discussed as effective approaches to improved sensitivity. For intact proteins, some sample pretreatment (depletion/enrichment) strategy was required to eliminate major matrix proteins. The high duty cycle of HRMS allows for detection of multiple tryptic peptides, which then allows post-acquisition identification of the most sensitive and selective fragment. Lindsay King (Pfizer, CT, USA) presented design considerations for LBAs that are used in early stages of a program to better understand PK, PD and immunogenicity of drug candidates. Emphasis was on assays to support ADC programs. Two different LBAs to measure ADCs were described. The advantages and disadvantages of each assay were compared as well as the utility of data obtained from each assay. The need to understand the influence of soluble or shed antigen on an LBA was discussed. If target concentrations are dynamic, the utility of an assay that measures both target-bound (total) and unbound (free) drug was highlighted. Incorporation of immunogenicity assessments was useful to better understand anomalous PK even during early stages of a program. Vimal Patel (Amgen, CA, USA) presented operational advantages and challenges of implementing automation in a bioanalytical laboratory. Multiple automated liquid handlers, their ease of use and programming were discussed. Generic scripts, once established, allow improved efficiency, data quality, pass-rate and reproducibility with minimum user input. A case study was presented showing integration of an automated microfluidics platform, a pipetting wizard and a laboratory information management system. A modern off-the-shelf pipetting wizard with a wide range of pipetting protocols and a laboratory information management system interface was adapted to fit sample pipetting and dilutions. Integration of modular applications of existing protocols and interfaces allowed for easy implementation with minimal resources. future science group
Conference Report | News & Analysis The Thursday morning plenary session on the final day featured four talks focusing on timely topics in bioanalysis. Rebbeca Sendak (Genzyme, MA, USA) discussed biosimilars and the challenges presented in order to analytically characterize complex biomolecules and demonstrate biosimilarity from a chemistry, manufacturing and control perspective. Through several examples she showed there is a spectrum of complexity with biomolecules making a single analytical approach impossible. The more complex the biomolecule the less one’s ability to fully perform analytical characterization. Analytical characterization of biosimilars requires a significant amount of planning and investment. Sendak discussed the bioanalytical challenges that biosimilars pose not only for PK analysis but also with evaluating immunogenicity. Surinder Kaur (Genentech, CA, USA) presented a very detailed overview of the analytical considerations for ADCs. ADCs have complex structures that require several methodologies and strategies in order to characterize the PK of the different components. She emphasized that success is highly dependent on extensive collaboration between the small- and large-molecule groups through the integration of ligand binding, LC–MS/MS and hybrid assay strategies and examples were presented. ADCs can also elicit an immune response that presents an analytical challenge because any of the component – antibody, linker and cytotoxic drug – may result in the generation of the antitherapeutic antibody response. Surinder emphasized that it is important to understand ADC structures in vivo to ensure appropriate assays are developed to understand PK and immunogenicity of these drug candidates. Jim Vrbanac (MPI Research, MI, USA) discussed the metabolites in safety testing guidelines as they apply to preclinical and clinical studies and described how high-resolution isotope-dilution MS (HRIDMS) can be a powerful tool in understanding the metabolism of drug candidates early in development. He discussed the use of isotopedilution MS as a precise and accurate technique and the use of HRMS versus MS/MS. Vrbanac presented several HRID-MS test experiments that demonstrated the utility of the technique. Due to recent advances in cell and tissue technologies the use of HRID-MS will continue to evolve. Peter van Amsterdam (Abbott, the Netherlands) presented a thorough comparison of the many regulatory guidelines focusing on bioanalysis. Started with a historical perspective he went on to show where the guidelines future science group
from the European Medicines Agency, FDA, ANVISA (Brazil), and MHLW (Japan) are harmonized and where they differ. His conclusion was that a bioanalytical group performing regulated analysis can begin with the European Medicines Agency guidelines as a base and make modifications from there. van Amsterdam gave a very extensive reference section at the end of his talk that would be useful to anyone delving into regulated bioanalysis. New afternoon sessions Because of the change in venue and shorter schedule, traditional afternoon workshops were replaced with additional plenary sessions. Tuesday afternoon’s program focused on outsourcing issues in bioanalysis from different segments of the pharmaceutical industry. Kenneth Ruterbories (Lilly, IN, USA) provided an overview of his company’s evolution from a fully integrated pharmaceutical company (everything in-house) to a fully integrated pharmaceutical network (a blend of in-house and outsourced analysis). Bioanalysis was identified as essential but not a core function and has evolved be a fully outsourced function. This evolution over 15 years resulted in reductions in instruments and personnel. Ruterbories reviewed the approach to selecting and narrowing down vendors to a small preferred list. The expectations for providers have evolved and these expectations have increased. He hopes to see an evolution in the way methods are developed; using good science but keep the methods simple. He expects CROs to be at the forefront of technology in areas such as eDocumentation and accurate quantification. He concluded that “outsourcing is an investment.” Brian O’Dwyer’s (ICON Development Solutions, NJ, USA) presented the CRO perspective and highlighted the challenges and evolving technologies. He focused on different aspects of managing partnerships with pharmaceutical companies and talked about the various challenges a CRO faces; it is a balance between meeting client’s expectations for a very high level of technical service and very slim operating margins. CROs need to be at the forefront of technology and new techniques while still maintaining profitability. O’Dwyer reported profitability data comparing eight Pharma companies and four CROs demonstrating that there were margins of 22 and 4%, respectively. However, the upside includes a large and growing percentage of bioanalysis work being outsourced. Closer partnerships between CROs and pharmaceutical companies can provide improved service and www.future-science.com
2603
News & Analysis | Conference Report cost savings to the benefit of both parties. Wei Zhou (Novartis, MA, USA) reviewed the challenges of outsourcing offshore. She gave an overview of her company’s philosophy of outsourcing. Novartis has also consolidated their bioanalytical units, but still keeps most work in-house through early-stage development. At that point they use a small number of vendors that have been carefully selected. For GLP studies they keep most studies in North America and Europe due to regulatory and logistical advantages. The cost at these sites is greater, but has decreased in recent years. CROs in China and India are used primarily for cost savings. The performance tends to be similar to other CROs around the world; however, the prices are rising and there are other disadvantages. Challenges in China include language, export of samples and time zones difference; with India there were recent changes in their import laws; and for Jordan and Russia it is primary a lack of familiarity. The Wednesday afternoon plenary session focused on prediction and prevention of immunogenicity. Crystal Sung (Sanofi, MA, USA) discussed the concept of immune tolerance induction (ITI) as it applies to drug development and patient treatment. She described the potential consequences of antidrug antibody formation, including its impact on PK, efficacy and safety. The concept and application of ITI was then discussed in the context of the enzyme replacement therapy used to treat Pompe Disease, a rare and often fatal disease caused by a deficiency of lysosomal acid a-glucosidase. While treatment for Pompe Disease exists, effectiveness is impacted by a high incidence of antidrug antibody formation in certain patient genotypes (e.g., crossreactive immunologic material – in infants). Various approaches to ITI were reviewed and clinical data were presented demonstrating how both prophylactic and therapeutic regimens with immune modulators can be used to obtain long-lived tolerance in Pompe Disease infants. Sung stressed that investigation into the mechanism of these responses is warranted so that translation to treatments for other life-threatening diseases can be understood. Bonita Rup (Pfizer, MA, USA) followed with a discussion on the state of the science for immunogenicity prediction, discussing some of the factors that influence immune responses to biotherapeutics proteins. These included product-related factors (e.g., sequence, aggregation, and post-translational modifications), patient influence (e.g., disease state, genetic factors and immune status) and treatment factor (e.g., regimen, route and co-medications). Rup discussed 2604
Bioanalysis (2013) 5(21)
the array of tools currently available for predicting immunogenicity prior to first-in-human trials, including: in silico T-cell epitope analysis, in vitro HLA binding assays, ex vivo dendritic cell T-cell activation assays, the artificial lymph node, and the concept of a ‘humanized mouse’ model. She described the advantages, limitations and future prospects of each of these tools with an emphasis on applicability. She concluded by emphasizing that many of these tools are in their infancy and require standardization and additional learning. The second part of Wednesday afternoon was the analytical investigators forum, which provided a platform for scientists to present research building on the major themes of the conference. Joy Hoesing (PRA, KS, USA) presented a case study investigation of whole blood sample collection and handling stability. Industry guidance indicates the necessity for evaluating sample collection and handling stability; however, there is no specific guidance on how the stability assessment should be conducted. Two specific whole blood sample collection and handling examples were covered in the presentation. The first case study demonstrated the possible impact of anticoagulant selection on analyte conversion, both in fortified whole blood samples and incurred samples. The second case study investigated whole blood stability in the presence of additional metabolites with results indicating the importance of conversion considerations in relation to both incubation time and temperature. Jonathan Haulenbeek (Bristol-Myers Squibb, NJ, USA) gave an overview of the importance of characterizing critical reagents for LBAs. Detailed attention was paid to defining the molar incorporation ratio of labeled critical reagents. A general process for labeling and subsequent analysis was given. Mass spectral techniques for incorporation ratio were the focus of development and late-phase case studies. The cases studies showed that analysis by MALDI-TOF-MS had aided assay optimization and trouble shooting. Detailed characterization of labeled reagents was shown to make method development and method transfer more efficient. The presentations initiated a lot of interest and discussions among the conference attendees. Opportunities for interacting & networking On Tuesday afternoon, following the plenary session, there were informal roundtables held on a variety of topics. These discussions and facilitators covered: future science group
Conference Report | News & Analysis Best practices of discovery bioanalysis: balancing data quality and productivity (Stacy Ho, Sanofi, MA, USA);
resulted in more interaction and greater camaraderie between participants than in previous years at the resort.
Personal skills development: managing through change in the laboratory (Randall H Guthrie, RHG & Associates, LLC, WI, USA);
Conclusion Due to the informal, noncommercial and improved conference setting, attendees have more time to meet each other, discuss their organizations, practices and the science than they would experience at large international conventions. Feedback was very positive and showed a significant improvement in evaluation questions related to the conference facility, food service and guest rooms. Next year’s conference, which is currently being developed, will follow a similar format at the same conference facility and will be posted on the University’s website when finished [2].
n
n
Dealing with daily bioanalytical laboratory problems (Chad Briscoe, PRA, KS, USA);
n
DBS and microsampling: what does the future hold? (Douglas J Turk, Lambda Therapeutic Research Inc., Canada);
n
Compliance challenges for the bioanalytical scientist (R John Stubbs, Stubbs and HenselPharma Consulting, LLC, PA, USA).
n
A major challenge with the new venue After 54 years of presenting Land O’Lakes Conferences at remote resorts, the major challenge facing the conference organizers was to create an interactive environment and keep people together with the distractions of an urban setting. This was accomplished by adding a new dinner presentation on the opening evening, better break service during the plenary sessions, roundtable discussions in a beer garden at a local brewery, and inclusion of gift cards for participants randomly selected at various times during the poster session. Initial impressions and conference evaluations support that these changes
Disclosure The views expressed in Naraharisetti’s presentation are those of the presenter and do not reflect official policy of the US FDA. No official endorsement by the FDA is intended or should be inferred.
Financial & competing interests disclosure The authors 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.
Websites 1
14th Annual Land O’Lakes Bioanalytical Conference – Emerging Challenges and Evolving Technologies for Bioanalysis. http://ce.pharmacy.wisc.edu/2013JulyLOLInfo
future science group
2
15th Annual Land O’Lakes Bioanalytical Conference (Madison, WI, USA) 21–24 July 2014 . http://ce.pharmacy.wisc.edu/2014JulyLOLInfo
www.future-science.com
2605
