An Avidin-Biotin Conjugation Technique for Presenting Target Antigens on Mycobacterium bovis BCG
Published: May 31, 2024
Abstract
Source: Liao, T. A., et al., Expression of Exogenous Antigens in the Mycobacterium bovis BCG Vaccine via Non-genetic Surface Decoration with the Avidin-biotin System. J. Vis. Exp. (2018).
The video demonstrates a technique for loading antigens on Mycobacterium bovis BCG to improve its immunogenic properties. The method uses the avidin-biotin system to coat the bacterial surface with exogenous antigens.
Protocol
1. Biotinylation of BCG Cell Surface Grow BCG in Middlebrook 7H9 broth with 10% OADC (oleic acid, albumin and dextrose solution) and 0.05% Tween 80 at 37 °C on a shaker platform at 50 rpm until OD = 0.5-1. NOTE: BCG is a biosafety level 2 pathogen and all experiments concerning BCG should be done in a biosafety cabinet with proper personal protective equipment. Wash 109 BCG 3 times with 500 µL of ice-cold endotoxin free PBS plus 0.1% Twee…
Divulgations
The authors have nothing to disclose.
Materials
| Endotoxin-free RPMI 1640 | StemCell Technologies | 36750 | |
| Sulfo-NHS SS biotin | Thermo Fisher | 21328 | |
| Middlebrook 7H9 broth | BD Diagnostic Systems | 271310 | |
| OADC | BD Diagnostic Systems | B11886 | |
| Tween 80 | Sigma-Aldrich | P1379 |
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Citer Cet Article
An Avidin-Biotin Conjugation Technique for Presenting Target Antigens on Mycobacterium bovis BCG. J. Vis. Exp. (Pending Publication), e22273, doi: (2024).