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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.
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.
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