Proteomic study via a non-gel based approach of meningococcal outer membrane vesicle vaccine obtained from strain CU385: A road map for discovering new antigens

Abstract

This work presents the results from a study of the protein composition of outer membrane vesicles from VA-MENGOC-BC (Finlay Institute, Cuba), an available vaccine against serogroup B Neisseria meningitidis. Proteins were identified by means of SCAPE, a 2DE-free method for proteome studies. More than one hundred proteins were detected by tandem liquid chromatographymass spectrometry analysis of fractions enriched in peptides devoid of histidine or arginine residues, providing a detailed description of the vaccine. A bioinformatic analysis of the identified components resulted in the identification of 31 outer membrane proteins and three conserved hypothetical proteins, allowing the cloning, expression, purification and immunological study of two of them (NMB0088 and NMB1796) as new antigens.

A deeper mining on the protein composition of VA-MENGOC-BC®: An OMV-based vaccine againstN. meningitidisserogroup B and C

“…This represents more than twice the individual contributions of any of the methods previously reported by our group. [19][20][21][22] One hundred and twelve proteins were identified in both incubation buffers ( Fig. 3E, and Supplementary Material 4), which is a cross confirmation of the presence of these proteins in the OMV preparation.…”

“…This conventional approach did not yield new protein identification, considering the previous results of our group. [19][20][21][22] To extend the protein catalogue of this OMV preparation we used the general strategy (HPLL&RP(OH ¡ /H C )) shown in Fig. 2.…”

“…This method, based on dual fractionation (DF) of proteins and peptides by polyacrylamide gel electrophoresis (PAGE), enabled the identification of 97 different proteins. 20 A gel-free method called SCAPE (Selective CApture of PEptides) 21 developed for 2DE-free proteome studies, was also applied to the characterization of this OMV preparation 22 and 106 non-redundant proteins were identified, including thirtyone, which were annotated or predicted to be membrane proteins. 22 Overlapping the information obtained in terms of protein identification, after applying the 2DE, DF-PAGE and SCAPE methods to the characterization of the OMV preparation, a total of 160 proteins were identified in this preparation.…”

Quantitative Proteomics of the Neisseria Gonorrhoeae Cell Envelope and Membrane Vesicles for the Discovery of Potential Therapeutic Targets

“…Sorting of these proteins into MVs was established in N. meningitidis and N. lactamica (75,76,78,79,86). One of the more fascinating features of a major GC porin, PorB (65) is its translocation from bacterial outer membrane and insertion into the membranes of eukaryotic cell organelles (87).…”

Charge state-selective separation of peptides by reversible modification of amino groups and strong cation-exchange chromatography: Evaluation in proteomic studies using peptide-centric database searches

“…Our group has reported several selective isolation methods based on chemical modification of peptide primary amino groups [8][9][10][11][12]. In this work, an integrating approach of these methods into a new separation scheme was developed.…”