Titlebook: Methods for Fragments Screening Using Surface Plasmon Resonance; Sameer Mahmood Zaheer,Ramachandraiah Gosu Book 2021 The Editor(s) (if app

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Preparation of Protein with Histidine Tag for Amine Couplingg with a relevant tag is critical. These tags can be used for ligating the protein onto the sensor with specific sensor chemistry. This chapter will outline the preparation of histidine tagged protein and can be used for the amine coupling technique.

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Protein Stability Using Thermal Shift Assay (TSA): pH Toleranceto the sensors, protein of interest need to be evaluated for pH tolerability and stability. Buffer mismatch is an important aspect of SPR assay due to its high sensitivity for small changes. Protein storage buffer is relatively different from immobilization and running buffer. To perform amine coupl

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Protein Stability Using Thermal Shift Assay (TSA): DMSO Toleranceto the sensors, protein of interest need to be evaluated for DMSO tolerability and stability. Buffer composition is an important aspect of SPR assay due to its high sensitivity for small changes in refractive indices. DMSO is one of the major factors that influence buffer mismatch leading to failure

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Selection and Identification of Fragment Librarys essential for starting fragment-based drug discovery efforts as multiple libraries are available commercially built on different clustering criteria, which are redundant to an extent. Although many criteria can be applied to select a diverse set of fragments toward specific targets, we describe ap

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Preparation of SPR Sensorstrument. Improper sensitization or activation of biosensors can lead to nonuniform ligation of ligands, which result in variations between triplicates during SPR assays. Inadequate activation is also responsible for the ligand drift post immobilization. This chapter provides information to prepare

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