Extraction of Membrane Components from Neisseria gonorrhoeae Using Catanionic Surfactant Vesicles : A New Approach for the Study of Bacterial Surface Molecules

Identification of antigens is important for vaccine production. We tested extraction protocols using cetyltrimethylammonium tosylate (CTAT) and sodium dodecylbenzenesulfonate (SDBS) to formulate surfactant vesicles (SVs) containing components from Neisseria gonorrhoeae. Carbohydrate and protein assays demonstrated that protein and carbohydrates were incorporated into the vesicle leaflet. Depending on the extraction protocol utilized, 100-400 µg of protein/mL of SVs solution was obtained. Gel electrophoresis followed by silver staining demonstrated that SV extracts contained lipooligosaccharide and a subset of bacterial proteins and lipoproteins. Western blotting and mass spectral analysis indicated that the majority of the proteins were derived from the outer membrane. Mass spectrometric and bioinformatics analysis of SVs identified 29 membrane proteins, including porin and opacity-associated protein. Proteins embedded in the SVs leaflet could be degraded by the addition of trypsin or proteinase K. Our data showed that the incorporation of CTAT and SDBS into vesicles eliminated their toxicity as measured by a THP-1 killing assay. Incorporation of gonococcal cell surface components into SVs reduced toxicity as compared to the whole cell extracts, as measured by cytokine induction, while retaining the immunogenicity. This process constitutes a general method for extracting bacterial surface components and identification of antigens that might be included in vaccines.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:12

Enthalten in:

Pharmaceutics - 12(2020), 9 vom: 20. Aug.

Sprache:

Englisch

Beteiligte Personen:

Stein, Daniel C [VerfasserIn]
H Stocker, Lenea [VerfasserIn]
Powell, Abigail E [VerfasserIn]
Kebede, Salsawi [VerfasserIn]
Watts, David [VerfasserIn]
Williams, Emma [VerfasserIn]
Soto, Nicholas [VerfasserIn]
Dhabaria, Avantika [VerfasserIn]
Fenselau, Catherine [VerfasserIn]
Ganapati, Shweta [VerfasserIn]
DeShong, Philip [VerfasserIn]

Links:

Volltext

Themen:

Antigen
Catanionic surfactant vesicles
Journal Article
Mass spectral analysis
Neisseria gonorrhoeae
Vaccine

Anmerkungen:

Date Revised 11.12.2021

published: Electronic

Citation Status PubMed-not-MEDLINE

doi:

10.3390/pharmaceutics12090787

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM313989532