Chapter 1. Introduction to Surface Plasmon Resonance -- Chapter 2. Principle of Surface Plasmon Resonance (OneStep) -- Chapter 3. Preparation of protein with Histidine Tag for Amine coupling -- Chapter 4. Preparation of protein with Avidin Tag for Biotin-based Capture -- Chapter 5. Protein stability using Thermal Shift Assay (TSA) - pH Tolerance -- Chapter 6. Protein stability using Thermal Shift Assay (TSA) - DMSO Tolerance -- Chapter 7. Selection and Identification of Fragment library -- Chapter 8. Preparation of Sensor -- Chapter 9. Optimization and validation of Amine coupling of Ligand -- Chapter 10. Optimization and Validation of Strepavidin/Biotin-based capture of Ligand -- Chapter 11. Preparation of Fragments for Screening in SPR -- Chapter 12. Screening of Fragments in SPR -- Chapter 13. Identification of Fragment Hits -- Chapter 14. Data Analysis and Confirmation of Hits.

This volume describes methods and protocols for the fragment-based screening of proteins using Surface Plasmon Resonance (SPR). The initial chapter of the book discusses the principle of SPR for the identification of biomolecular interactions, while the subsequent chapters introduce methods for labelling proteins with different tags including, histidine and biotin tags. It also discusses techniques and factors that affect the amine and biotin-streptavidin coupling and methods to optimize the interactions. Next, it describes fragment preparation for screening in SPR and presents methods to calculate equilibrium dissociation constant (KD) and ligand efficiency (LE). It reviews techniques of next-generation injections that improve the efficiency of the characterization process over traditional SPR by determining the kinetics and affinity in a single step. Finally, the book elucidates a comprehensive yet representative description of challenges associated with the molecular interaction of proteins using SPR.