Methods for Fragments Screening Using Surface Plasmon Resonance

This book provides methods and protocols for fragment-based screening of proteins using Surface Plasmon Resonance (SPR). It discusses the principle of SPR, labelling techniques, fragment preparation, equilibrium dissociation constant calculation, and next-generation injections. It also highlights challenges associated with protein molecular interactions using SPR.

Format: Paperback / softback
Length: 113 pages
Publication date: 17 September 2022
Publisher: Springer Verlag, Singapore

This comprehensive volume provides detailed methods and protocols for conducting fragment-based screenings of proteins using Surface Plasmon Resonance (SPR). The book begins by delving into the fundamental principles of SPR, elucidating its role in identifying biomolecular interactions. Subsequently, it explores various strategies for labeling proteins with different tags, such as histidine and biotin tags, which are essential for subsequent SPR analyses. The chapters also discuss techniques and factors that influence the amine and biotin-streptavidin coupling, emphasizing the importance of optimizing these interactions for accurate measurements.

The volume then proceeds to describe fragment preparation for screening in SPR, including methods to calculate equilibrium dissociation constant (KD) and ligand efficiency (LE). It reviews advanced injection techniques, such as next-generation injections, which enable the simultaneous determination of kinetics and affinity in a single step, greatly enhancing the efficiency and accuracy of the characterization process compared to traditional SPR.

Furthermore, the book offers a comprehensive yet representative account of the challenges associated with the molecular interaction of proteins using SPR. It discusses the limitations of SPR in detecting weak interactions, the influence of protein conformations on binding behavior, and the need for proper experimental design and data analysis to obtain reliable and meaningful results.

By presenting these comprehensive methods and protocols, this volume serves as a valuable resource for researchers and scientists engaged in fragment-based screening, protein labeling, and SPR-based molecular interaction studies. It provides a solid foundation for understanding the principles, techniques, and considerations involved in these fields, enabling researchers to advance their understanding of protein function and interactions.

Weight: 215g
Dimension: 235 x 155 (mm)
ISBN-13: 9789811615382
Edition number: 1st ed. 2021