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The first book to offer a blueprint for overcoming the challenges to successfully quantifying biomarkers in living organisms
The demand among scientists and clinicians for targeted quantitation experiments has experienced explosive growth in recent years. While there are a few books dedicated to bioanalysis and biomarkers in general, until now there were none devoted exclusively to addressing critical issues surrounding this area of intense research. Target Biomarker Quantitation by LC-MS provides a detailed blueprint for quantifying biomarkers in biological systems. It uses numerous real-world cases to exemplify key concepts, all of which were carefully selected and presented so as to allow the concepts they embody to be easily expanded to future applications, including new biomarker development.
Target Biomarker Quantitation by LC-MS primarily focuses on the assay establishment for biomarker quantitation--a critical issue rarely treated in depth. It offers comprehensive coverage of three core areas of biomarker assay establishment: the relationship between the measured biomarkers and their intended usage; contemporary regulatory requirements for biomarker assays (a thorough understanding of which is essential to producing a successful and defendable submission); and the technical challenges of analyzing biomarkers produced inside a living organism or cell.
* Covers the theory of and applications for state-of-the-art mass spectrometry and chromatography and their applications in biomarker analysis
* Features real-life examples illustrating the challenges involved in target biomarker quantitation and the innovative approaches which have been used to overcome those challenges
* Addresses potential obstacles to obtain effective biomarker level and data interpretation, such as specificity establishment and sample collection
* Outlines a tiered approach and fit-for-purpose assay protocol for target biomarker quantitation
* Highlights the current state of the biomarker regulatory environment and protocol standards
Target Biomarker Quantitation by LC-MS is a valuable resource for bioanalytical scientists, drug metabolism and pharmacokinetics scientists, clinical scientists, analytical chemists, and others for whom biomarker quantitation is an important tool of the trade. It also functions as an excellent text for graduate courses in pharmaceutical, biochemistry and chemistry.
List of contents
List of Contributors xv
Preface xix
Abbreviations xxiii
Part I Overview 1
1 Overview of Targeted Quantitation of Biomarkers and Its Applications 3
Naidong Weng
1.1 Introduction 3
1.2 Biomarker Definition 4
1.3 Current Challenges of a Biomarker 5
1.4 Biomarker Validation Process 6
1.5 Current Regulatory Requirement for Target Biomarker Quantitation 6
1.6 Challenges of Biomarker Quantitation 7
1.7 Current Technologies for Biomarker Quantitation 8
1.7.1 LC-MS 8
1.7.2 GC-MS 8
1.7.3 Ligand?]Binding Assay 9
1.7.4 Flow Cytometry 9
1.7.5 Quantitative PCR (qPCR) 9
1.8 Current Biomarker Quantitation Applications 9
1.8.1 Protein Biomarkers 9
1.8.2 Peptide Biomarkers 10
1.8.3 RNA Biomarkers 11
1.8.4 Nucleotide Biomarkers 11
1.8.5 Small Molecule Biomarkers 11
1.9 Conclusion and Future Perspective 12
References 13
2 Translational Application of Biomarkers 17
Ray Bakhtiar
2.1 Introduction 17
2.2 Translational Medicine 17
2.3 Biomarkers 18
2.4 Biomarker Categories 18
2.5 Neurobiological Disorders 21
2.6 Cardiovascular Disorders 22
2.7 Chronic Obstructive Pulmonary Disease 23
2.8 Oncology 24
2.9 Biomarker Measurements and Regulatory Considerations 26
2.10 Conclusions 27
References 29
3 Current Regulatory Guidance Pertaining Biomarker Assay Establishment and Industrial Practice of Fit?]for?]Purpose and Tiered Approach 35
Naidong Weng
3.1 Introduction 35
3.2 Current Regulatory Guidance and Interpretation 36
3.3 Current Industrial Discussion and Recommendations 37
3.4 Considerations for Assay Validation and Sample Analysis 39
3.4.1 Sensitivity 40
3.4.2 Specificity and Selectivity 40
3.4.3 Matrix Effects and Sample Variables 40
3.4.3.1 Authentic Analyte/Authentic Matrix Approach 40
3.4.3.2 Surrogate Analyte/Authentic Matrix Approach 40
3.4.3.3 Authentic Analyte/Surrogate Matrix Approach 40
3.4.4 Accuracy/Precision 40
3.4.5 Stability 41
3.4.6 Sample Analysis Consideration 41
3.5 Examples of Fit?]for?]Purpose and Tiered Approach 41
3.5.1 Relative Quantification of Glyco?]isoforms of Intact Apolipoprotein C3 in Human Plasma by LC?]HRMS 41
3.5.2 Quantification of 4ß?]Hydroxycholesterol Endogenous Biomarker for CYP3A4 Activity in Plasma Samples 41
3.5.3 Quantitation of Leukotriene B4 in Human Sputum as a Biomarker Using UPLC-MS/MS 42
3.6 Conclusion 42
References 42
4 Modern Liquid Chromatography and Mass Spectrometry for Targeted Biomarker Quantitation 45
Wenying Jian
4.1 Introduction 45
4.2 Liquid Chromatography 45
4.2.1 Importance of Separation 45
4.2.2 Basic Principle of LC 47
4.2.3 Major Modes of LC Used for Targeted Biomarker Quantitation 47
4.2.4 Modern LC Technologies 49
4.2.4.1 HPLC and UHPLC 49
4.2.4.2 Miniaturized Column LC 50
4.2.4.3 2D?]LC 51
4.3 Mass Spectrometry 51
4.3.1 Major Types of MS Used for Targeted Biomarker Quantitation 51
4.3.2 Ionization Techniques 54
4.3.3 Ion Mobility 54
4.3.4 Fragmentation Mode 55
4.3.5 Emerging MS Techniques 56
4.3.5.1 MS Imaging 56
4.3.5.2 Other Surface Analysis MS Techniques 58
4.4 Summary and Future Perspectives 58
Referenc
About the author
Naidong Weng, PhD, is Scientific Director, Janssen Fellow, and Head of Bioanalytical Chemistry and Pharmacokinetics within Department of Pharmacokinetics, Dynamics and Metabolism at US East Coast, Janssen Research & Development, Johnson and Johnson. He has over 25 years of experiences on quantitative bioanalysis. His research interest includes using HILIC-MS/MS for quantitation of highly polar analytes as well as chiral analysis. He has published more than 110 journal papers and book chapters.
Wenying Jian, PhD, a Senior Principal Scientist of Bioanalytical Chemistry and Pharmacokinetics within Department of Pharmacokinetics, Dynamics and Metabolism at US East Coast, Janssen Research & Development, Johnson and Johnson. Her research experience and interest center on application of advanced LC-MS methodologies in detection, identification, and quantitation of endogenous molecules, drugs and their metabolites, including small and large molecules, and in complicated biological matrices. She has published more than 40 journal papers and book chapters.
Summary
The first book to offer a blueprint for overcoming the challenges to successfully quantifying biomarkers in living organisms The demand among scientists and clinicians for targeted quantitation experiments has experienced explosive growth in recent years.
