Development of SCADA-based automation of affinity chromatography for high-throughput two-step protein purification.

摘要

Structural or functional protein studies require large amounts of highly pure proteins. To achieve this manually requires a lot of time and effort. Manual procedures are also inefficient and error-prone, and the protein yield is limited. Semi-Automated Protein Purification Machine for Affinity Chromatography (SAPPMAC) has been designed to reduce the time and efforts required for protein purification, while increasing the yield through parallel processing. A two-step procedure, utilizing the benefits of Immobilized Metal-Ion Affinity Chromatography (IMAC), which exploits the affinities of certain tags, like histidine, for metal ions, has been chosen as the protocol for automation. A GUI for representing the real-time status of the process has been built. The GUI also provides two modes of control for controlling the SAPPMAC.;Flexibility and scalability provide the most important features of the SAPPMAC. The instrument flexibility is provided by user-defined configurations and more such units can be operated in parallel to make the instrument scalable for high throughputs. The GUI gives simple operation through process visualization and, automatic mode and manual mode of control. The semi-automated approach provides cost efficient operation, low maintenance and increased labor efficiency. These features make SAPPMAC a utility machine for laboratory operations and parallel purification runs when large amounts of highly pure proteins are required.