Preformulation studies with the Escherichia coli double mutant heat-labile toxin adjuvant for use in an oral vaccine.
J Immunol Methods
; 451: 83-89, 2017 12.
Article
en En
| MEDLINE
| ID: mdl-28939395
ABSTRACT
Double mutant heat-labile toxin (dmLT) is a promising adjuvant for oral vaccine administration. The aims of our study were to develop sensitive methods to detect low concentrations of dmLT and to use the assays in preformulation studies to determine whether dmLT remains stable under conditions encountered by an oral vaccine. We developed a sandwich ELISA specific for intact dmLT and a sensitive SDS-PAGE densitometry method, and tested stability of dmLT in glass and plastic containers, in saliva, at the pH of stomach fluid, and in high-osmolarity buffers. The developed ELISA has a quantification range of 62.5 to 0.9ng/mL and lower limit of detection of 0.3ng/mL; the limit of quantification of the SDS-PAGE is 10µg/mL. This work demonstrates the application of dmLT assays in preformulation studies to development of an oral vaccine containing dmLT. Assays reported here will facilitate the understanding and use of dmLT as an adjuvant.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Toxinas Bacterianas
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Ensayo de Inmunoadsorción Enzimática
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Adyuvantes Inmunológicos
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Proteínas de Escherichia coli
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Electroforesis en Gel de Poliacrilamida
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Enterotoxinas
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Mutación
Idioma:
En
Revista:
J Immunol Methods
Año:
2017
Tipo del documento:
Article