Polymorphic distribution of proteins in solution by mass spectrometry: The analysis of insulin analogues.

Autor: Fávero-Retto MP; Federal University of Rio de Janeiro - UFRJ, Av. Carlos Chagas Filho 373, CCS, Bss24, Ilha do Fundão, 21941-902, Rio de Janeiro, RJ, Brazil; Brazilian National Cancer Institute (INCA), 20230-014, Rio de Janeiro, RJ, Brazil., Guerreiro LH; Federal University of Rio de Janeiro - UFRJ, Av. Carlos Chagas Filho 373, CCS, Bss24, Ilha do Fundão, 21941-902, Rio de Janeiro, RJ, Brazil; Department of Chemistry, Institute of Exact Sciences, Rural Federal University of Rio de Janeiro - UFRRJ, Rodovia BR 465, km 7, CEP: 23890-000, Seropédica, RJ, Brazil., Pessanha CM; Federal University of Rio de Janeiro - UFRJ, Av. Carlos Chagas Filho 373, CCS, Bss24, Ilha do Fundão, 21941-902, Rio de Janeiro, RJ, Brazil., Palmieri LC; Federal University of Rio de Janeiro - UFRJ, Av. Carlos Chagas Filho 373, CCS, Bss24, Ilha do Fundão, 21941-902, Rio de Janeiro, RJ, Brazil., Lima LMTR; Federal University of Rio de Janeiro - UFRJ, Av. Carlos Chagas Filho 373, CCS, Bss24, Ilha do Fundão, 21941-902, Rio de Janeiro, RJ, Brazil; Laboratory for Macromolecules (LAMAC-DIMAV), Brazilian National Institute of Metrology, Quality and Technology - INMETRO, Av. N. Sa. das Graças, 50 - Xerém, Duque de Caxias-RJ, 25250-020, Rio de Janeiro, Brazil; National Institute of Science and Technology for Structural Biology and Bioimaging (INBEB-INCT), Federal University of Rio de Janeiro, Rio de Janeiro, 21941-590, Brazil. Electronic address: Mauricio@pharma.ufrj.br.
Jazyk: angličtina
Zdroj: Biologicals : journal of the International Association of Biological Standardization [Biologicals] 2017 Jan; Vol. 45, pp. 69-77. Date of Electronic Publication: 2016 Oct 27.
DOI: 10.1016/j.biologicals.2016.09.011
Abstrakt: The characterization of conformational and oligomeric distribution of proteins is of paramount importance for the understanding of the correlation between structure and function. Among the bioanalytical approaches currently available, the electrospray ionization-mass spectrometry (ESI-MS) coupled to ion mobility spectrometry (IMS) is the best suited for high resolution identification with high sensitivity, allowing the in situ separation of oligomeric and conformational species. We tested the performance of the ESI-MS technique along with the IMS separation approach on a broad variety of insulin and insulin analogues with distinct oligomeric distribution pattern. The measurement of commercial insulin allowed the identification of species ranging from monomers to hexamers and their complexes with zinc ions. Dissimilar distribution profile for regular insulin as a function of formulation component and among the insulin analogues were observed by ESI-IMS-MS but not by ESI-MS along, crystallographic assays or size-exclusion chromatography. These data suggest the additional suitability of ESI-IMS-MS in conformational and oligomeric profiling of biomacromolecules and biopharmaceuticals. The easiness of the technique provides further motivation for its application in the characterization of both raw and formulated protein biopharmaceuticals in routine and comparability exercises.
(Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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