Meet SV's new Protein Production and Structure Core Facility
Florence Pojer and David Hacker at the PPSCF © Nik Papageorgiou/EPFL
EPFL's School of Life Sciences has inaugurated a new Protein Production and Structure Core Facility (PPSCF), which will provide high-quality support to ongoing research on and off campus.
The PPSCF is administrated by Dr Florence Pojer and Dr David Hacker, who will be joining their respective expertise in protein crystallography and production. As all of SV’s Core Facilities, the PPSCF will function as a powerful means of optimizing research efforts in and even outside of EPFL. Although logistically the PPSCF has already been instituted, it will formally begin operations in late 2017.
We caught up with Drs Pojer and Hacker to ask them about the new platform.
What scientific questions does the PPSCF answer?What is its mission?
Florence Pojer: Our mission is to provide expertise and instrumentation in the field of protein production, purification, and structural characterization to the EPFL community as well as outside the campus. We are a team of specialists trained in all the different aspects of proteins going from the production and purification to biophysical and structural characterization.
David Hacker: From the protein production side, our mission is to produce recombinant proteins from microbial (E. coli) and animal cells. The latter category includes human embryonic kidney (HEK293), Chinese hamster ovary (CHO), and DrosophilaSchneider (S2) cells. We also produce monoclonal antibodies from hybridoma cell lines, and we grow other cell lines for projects concerning genomics and proteomics. We also offer services in protein purification.
How does the underlying technology work? What principles does it take advantage of?
FP: Our technologies rely on state of the art instrumentations located at one place in EPFL’s AI building, where the users can freely come to use them or to interact with other scientists. The PPSCF facility is meant to be a hub for sharing projects, ideas and expertise.
The strength of the facility lies in close interactions with outside institutes to benefit from a specialized set-up. For example, our users have the opportunity to regularly send protein crystal samples to Swiss Light Source at Villigen allowing them to get high-resolution 3D structures of their desired target protein.
DH: For the cultivation of animal cells in serum-free suspension cultures we rely on “low-tech” orbitally shaken bioreactors. Essentially, we use cylindrical and square-shaped glass bottles for the cultivation of cells from volumes of 100 mL to 10 L. For smaller volumes we use TubeSpin50 tubes. All of these containers are agitated in CO2 incubators by orbital shaking.
This technology was developed at EPFL in the Laboratory of Cellular Biotechnology (LBTC) of Professor Florian Wurm – now retired – in collaboration with Kühner AG in Basel, Techno Plastic Products in Trasadingen, and ExcellGene SA in Monthey. The LBTC also developed methods for the efficient transfection of HEK293, CHO, and S2 cells in serum-free suspension cultures. We use this technology to produce recombinant proteins transiently from these cell hosts.
In addition, the LBTC also developed an efficient method for the generation of stable HEK293, CHO, and S2 cell lines (monoclonal) and cell pools (polyclonal) for recombinant protein production using the PiggyBac Transposon System for gene delivery.
Who will be using the Facility? Will it be restricted to EPFL or also external researchers?
FP: The facility is of course open to researchers outside EPFL, as long as EPFL labs, and in particular SV labs, are not left behind.
DH: In 2016 we worked with 43 different research labs, with 24 of them being from the EPFL SV, SB, and STI faculties. Most of the remaining clients were located at the universities of Lausanne and Geneva. We only work with academic labs and have no commercial clients.
What kinds of projects will the PPSCF support? What advantages will it offer them?
FP: The projects are very diverse but often therapeutically oriented. To get a fuller view, projects need nowadays to combine many techniques that are often challenging and not available in their own labs. So at the PPSCF we provide scientists with both the expertise and the instruments, to allow them to stay competitive and publish in high-impact factor journals.
DH: Our proteins are used for protein structure studies, for functional studies, for reagents (such as antibodies for flow cytometry), for therapeutic studies, and as growth factors for stem cell culture.
In many cases, the proteins are not available commercially or they are very expensive. This is the case for growth factors for stem cell culture, for example. Making these proteins in-house means a better use of financial resources for several labs at the EPFL.
How will PPSCF be structured?
FP: At the facility, we are a team of experts in different field of proteins. But an important aspect that needs to be mentioned is the interaction with EPFL research labs. Those interactions are crucial for both entities to stay competitive and up-to-date. One example is our close contact with Professor Matteo Dal Peraro’s lab, an expert in molecular dynamics of proteins, and Professor Hilal Lashuel’s lab, an expert in the chemical synthesis and biophysical characterization of proteins. We will greatly benefit from their expertise. Moreover, an interactive website will also be created to link scientists, expertise and instruments.
DH: While continuing to produce proteins for EPFL research groups, we will also increase our services in protein purification. In the past, we only offered a single step of purification by affinity chromatography; now we have the expertise and equipment to perform additional purification steps based on protein size or charge. Of course, we will maintain open-access equipment for researchers who want to do their own purifications.
As we did before, we will provide services and training for determining protein structure by X-ray diffraction and assist with NMR approaches. Plus, we will maintain several open-access instruments for the biophysical characterization of proteins. In short, we will be able to offer a full array of services from protein production to purification to characterization under one roof.
What is your vision for the PPSCF for the future?
FP: Researchers are often in “transition” at EPFL; coming here from different horizons for a couple of years. Facilities like PPSCF are “permanent” entities, so one of our goals is to bring researchers together. We want the PPSCF to be a hub for researchers to meet, interact, share, and form collaborations. To intensify collaboration between facilities and research labs, as we both try to reach the same goal of moving research forward.
DH: To answer this, I think we first have to see how the platform functions after the fusion of the two core facilities. The PECF will not move to the PPSCF space until the end of 2017. So until we spend some time together it is hard to judge how to think about the future.