Bacterial Culture Filtrate Protein Concentration using an Ultrafiltration Device
Published: November 30, 2023
Abstract
Source: Ryan, J. M., et al. Mycobacterium tuberculosis Extracellular Vesicle Enrichment through Size Exclusion Chromatography. J. Vis. Exp. (183) (2022)
This video demonstrates the isolation and concentration of bacterial culture filtrate proteins, which include bacterial extracellular vesicles, using ultrafiltration. The concentrated extracellular vesicles serve as diagnostic markers for disease detection.
Protocol
1. Preparation of crude Mycobacterium tuberculosis (Mtb) extracellular vesicle (EV) concentrate NOTE: For detailed procedures on the cultivation of Mtb and preparation of culture filtrate protein (CFP). It is recommended that bacterial culture media is free from growth supplements with EV-containing or proteinaceous components, such as Oleic Albumin Dextrose Catalase (OADC), and detergents such as Tween. It is also recommended that the bacterial culture…
Disclosures
The authors have nothing to disclose.
Materials
| Automatic Fraction Collector | IZON Science | AFC-V1-USD | |
| Centricon Plus – 70 Centrifugal filter, 100 kDa cutoff | Millipore Sigma | UFC710008 | Ultrafiltration device used in step 1.1 |
| Phosphate-buffered Saline, 1X without calcium and magnesium | Corning | 21-040-CV | |
| Nitrocellulose membrane, Roll, 0.2 μm | BioRad | 1620112 |
Cite This Article
Bacterial Culture Filtrate Protein Concentration using an Ultrafiltration Device. J. Vis. Exp. (Pending Publication), e21874, doi: (2023).