Project Work Package 2


Empowering the service capacity for protein production in bacteria and in mammalian cells

Leader: Prof. Marco Fragai

  • Development of a platform for a high-throughput protein expression. (UNIFI-CERM)
  • Expanding the mammalian expression platform for in-cell NMR. (UNIFI-CERM)
  • Empowering Protein production services @CNR. (CNR-IC-Caserta)

In this work-package the isotopic labelled protein expression platform available at CERM/CIRMMP will be upgraded with an expanded set of equipment for chromatography to increase the lab throughput. Our chromatographic systems, based on ÄKTA technology, have to be upgraded with instruments able to satisfy different user requests.
Particularly, CERM laboratory will be equipped with a ÄKTA Pure L1, a highly versatile modular system, and the ÄKTA Pure M1 system, which includes a multiwavelength UV and visible spectrum detector from 190 to 700 nm (up to three wavelengths simultaneously) for proteins with different absorption profiles.
The new systems support a wide range of chromatography techniques and meet the automation requirements needed to deliver higher purity of samples. The possibility to configure the system with a wide range of options (multiple wavelengths detectors, air sensors, multiple pumps, etc..) further increase its capabilities depending on the user’s purification needs, in addition to a considerable time-saving due to the higher level of automation. The cited expected results will be easily measurable in terms of purity and stability grade of purified products and most importantly in terms of final protein yields.
Once delivered, the instruments will be installed by the provider, under the supervision of the technical staff dedicated to this activity. The scientific staff specifically hired for the purpose, will test the equipment on a set of protein constructs already available in the lab, for which protocols have been already developed.
Upon passing the initial tests, the equipment will be used by CERM/CIRMMP researchers and by the personnel specifically hired for the project to optimize the purification strategies currently used for various proteins, to evaluate the improvements in yield and throughput of the expression platform. These tests will result in a number of updated protocols and workflows. Finally, the platform will be made available to external users.