Data di Pubblicazione:
2012
Citazione:
Innovative high resolution mass spectrometry applications in drug discovery / G. Aldini. ((Intervento presentato al 6. convegno Nuove prospettive in chimica farmaceutica tenutosi a Riccione nel 2012.
Abstract:
In the past, the main applications of mass spectrometry in the pharmaceutical field
were mainly oriented towards structure elucidation and quantitative analysis of drug
and metabolites in biological matrices. Since the advent in the 90’s of soft ion sources
such as ESI and MALDI, allowing the analysis of biomacromolecules (biopolymers
such as DNA, proteins, peptides and sugars), MS has found growing applications in the
different steps of drug discovery and development processes, including target
identification, hit identification and lead identification and optimization.
In particular, ESI source is the workhorse for the mass spectrometric analysis of
noncovalent bound protein complexes since it allows intact weakly bound complexes to
be detected reflecting the nature of the interaction found in the condensed phase.
Several applications of such techniques have been reported, the most popular being
the identification of selective and high affinity ligands by fishing libraries or using
fragment based approaches. In particular, ligands are easily identified on the basis of
the MW of the protein adduct and their affinity established on the basis of their relative
abundance. The strength of this technique lies in its inherent sensitivity (picomolar
quantities of protein and ligand required), its speed (as low as 30 s for each analysis),
and its ability to simultaneously determine binding stoichiometry and dissociation
constants. A further improvement in studying non covalent interactions came about
more recently with the advent of nano-ESI source and high resolution MS analyzers
such as orbitrap. Several applications have been reported and we have successfully
applied this technique to screen targeted libraries against proteins [1,2] and nucleic
acid [3].
Moreover, the effect of ligand interaction on protein conformation can be evaluated by
studying the distribution of multicharged ions recorded in non denaturating conditions
as well as the protein aa involved in the ligand recognition by H/D exchange
experiments.
The most recent investigation indicates that MS is also an excellent method for
determining binding strength of protein-protein interactions within noncovalent
complexes. The substantial advantages of MS over other techniques (e.g., ITC) are
sensitivity and the ability to provide information on composition, stoichiometry and
subunit interactions of protein complexes.
References
[1] L. Regazzoni, L. Bertoletti, G. Vistoli et al., ChemMedChem, 2010, 5, 1015-1025
[2] L. Regazzoni, R. Colombo, L. Bertoletti et al., Anal Chim Acta, 2011
[3] F. Riccardi Sirtori, G. Aldini, M. Colombo et al., submitted
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
G. Aldini
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