Flow Cytometry in Drug Discovery and Development

This book covers the unique application of flow cytometry in drug discovery and development. The first section includes two introductory chapters, one on flow cytometry and one on biomarkers, as well as a chapter on recent advances in flow cytometry. The second section focuses on the unique challenges and added benefits associated with the use of flow cytometry in the drug development process. The third section contains a single chapter presenting an in depth discussion of validation considerations and regulatory compliance issues associated with drug development.

Preface xi

Foreword xiii

Acknowledgments xv

Contributors xvii

Part I Introduction 1
Philip Marder and Virginia Litwin

1 Introduction to Flow Cytometry 3
Elizabeth Raveche, Fatima Abbasi, Yao Yuan, Erica Salerno, Siddha Kasar, and Gerald E. Marti

2 Recent Advances in Flow Cytometry: Platforms, Tools, and Challenges for Data Analysis 23
Paul J. Smith, Roy Edward, and Rachel J. Errington

3 Introduction to Biomarkers 55
Ole Vesterqvist and Manjula P. Reddy

Part II Flow Cytometry In The Drug Development Process 69
Virginia Litwin and Philip Marder

4 HTS Flow Cytometry, Small-Molecule Discovery, and the NIH Molecular Libraries Initiative 71
Larry A. Sklar and Bruce S. Edwards

5 A Multiparameter Approach to Cell Cycle Analysis as a Standard Tool in Oncology Drug Discovery 99
Carmen Raventos-Suarez and Byron H. Long

6 Flow Cytometry in Preclinical Toxicology/Safety Assessment 123
David McFarland and Kristi R. Harkins

7 Use of Flow Cytometry to Study Drug Target Inhibition in Laboratory Animals and in Early-Phase Clinical Trials 151
David W. Hedley

8 CD4 T Cell Assessments in Evaluation of HIV Therapeutics 169
Thomas N. Denny, Raul Louzao, John Wong, and Brooke Walker

9 Monitoring the Cellular Components of the Immune System During Clinical Trials: A Translational Medicine Approach 189
Virginia Litwin and James Andahazy

10 Immunogenicity Testing Using Flow Cytometry 205
Denise M. O.Hara and Valerie Theobald

11 Pharmacokinetics by Flow Cytometry: Recommendations for Development and Validation of Flow Cytometric Method for Pharmacokinetic Studies 225
Yuanxin Xu and Susan M. Richards

Part III Validation And Regulatory Compliance 241
Virginia Litwin and Philip Marder

12 Regulatory Compliance and Method Validation 243
Carla G. Hill, Dianna Y. Wu, John Ferbas, Virginia Litwin, and Manjula P. Reddy

13 Instrument Validation for Regulated Studies 267
John Ferbas and Michelle J. Schroeder

Part IV Future Directions 279
Virginia Litwin and Philip Marder

14 Probability State Modeling: A New Paradigm for Cytometric Analysis 281
C. Bruce Bagwell

15 Phospho Flow Cytometry: Single-Cell Signaling Networks in Next-Generation Drug Discovery and Patient Stratification 303
Peter O. Krutzik, Sean C. Bendall, Matthew B. Hale, Jonathan M. Irish, and Garry P. Nolan

Index 335

"Flow Cytometry in Drug Discovery and Development is a monograph which comprehensively addresses the technical aspects of flow cytometry applied to drug development. Readers will learn how the unique benefits of this powerful analytical technique can be realized in the drug development process." (Analytical and Bioanalytical Chemistry, 29 July 2012)

"This book covers the unique application of flow cytometry in drug discovery and development." (Smart Grid, 15 February 2011) Virginia Litwin is a Principal Scientist at Covance Central Laboratory Services. After numerous years in the pharmaceutical/biotechnology industry, she has become a leader in the application and validation of flow cytometric methods in drug discovery and development. She founded the Flow Cytometry Subcommittee of the Ligand Binding Assay Bioanalytical Focus Group within the American Association of Pharmaceutical Scientists (AAPS). She is an active contributor to the International Clinical Cytometry Society (ICCS) and the International Society for Analytical Cytology (ISAC). She received her bachelor's from the University of California and her doctorate from the University of Iowa.

Philip Marder was internationally distinguished for his leadership in the application of flow cytometry in early drug discovery and development. He was a key contributor to the scientific community as an active member of ISAC and AAPS. After receiving both his bachelor's and master's degrees from The Ohio State University, he had a distinguished thirty year career at Eli Lilly and Company.

Overview of Flow Cytometry in Drug Discovery and Development

In this book, the unique application of flow cytometry in pharmaceutical discovery and development is presented. A basic explanation of the technology precedes a comprehensive review of specific examples of how the technology is utilized in drug development.

Readers of this book will learn how flow cytometry's use of multiparametric, high-speed measurements and manipulations of single cells adds valuable information and efficiencies for the discovery and development of new, better, and more affordable pharmaceuticals. It discusses validation and regulatory compliance issues as well as the use of cytometry in monitoring clinical diseases and evaluating therapies.

Divided into four parts, the book:

• Includes two introductory chapters, one on flow cytometry and one on biomarkers, as well as a chapter on recent advances in flow cytometry

• Focuses on the unique challenges and added benefits associated with the use of flow cytometry in the drug development process

• Presents an in-depth discussion of validation considerations and regulatory compliance issues associated with drug development

• Ends with two chapters addressing important future directions in the field of flow cytometry

Written by thought-leaders from academia and industry as well as flow cytometry users from the pharmaceutical sector, this comprehensive and straightforward text helps pharmaceutical researchers understand the potential of flow cytometry and, in turn, allows flow cytometrists to understand the needs of the pharmaceutical industry.