More for Less: Low-Cost, High-Yield Protein Deuteration for Neutron Crystallography
A team from Lund Protein Production Platform, Karolinska Institute, and the European Spallation Source have published an optimized approach to high-yield protein deuteration at low cost, for neutron protein crystallography. Read the full story here.
Baculovirus-driven protein expression in insect cells: A benchmarking study
Baculovirus-insect cell expression system has become one of the most widely used eukaryotic expression systems for heterologous protein production in many laboratories. To date, limited data has been presented in the literature to benchmark the protocols used for baculovirus vectors, and this study facilitates the selection of a system for optimal production of target proteins.
The benchmarking study was conducted by 13 Protein Production and Purification Partnership in Europe (P4EU) member facilities, including Lund Protein Production Platform. Taking advantage of the P4EU scientific network, a benchmarking initiative was designed to compare the diverse protocols established in thirteen individual laboratories. This study conducted during the last 4 years was published recently
Beamtime at BioMAX
LP3 has been granted beamtime at BioMAX (the X-ray macromolecular crystallography beamline of MAX IV). This allows users to have their crystals rapidly evaluated for diffraction and with the funding from BioCARE (a governmental supported strategic research area in cancer) the LP3 can also offer the possibility to process data and solve structures. Help in fishing and flash freezing crystals can be provided by LP3. In the spring of 2018, LP3 has beamtime at BioMAX on March& 23 and May 4.
New equipment for differential scanning fluorimetry (DSF)
A Prometheus NT.48 (nanotemper) is now available at LP3 for differential scanning fluorimetry (DSF) experiments. Users interested in running the instrument should contact maria [dot] gourdon [at] biol [dot] lu [dot] se.
Read more about the instrument here.
Protein Production Network Sweden launches new website
LP3 is one of several protein production facilities in Sweden that form the Protein Production Network Sweden (PPNS), each with its own specialties and focus. PPNS is an informal network formed in 2013 between protein production facilities at Swedish universities. The overall aim is to make competence and protein production methodologies available to the Swedish academic life science community and to make best use of our available resources. A link to the PPNS homepage and presentation, as well as contact details of the participating facilities can be found here.
Partnership between LP3 and DEMAX
In 2016, the DEuteration and MAcromolecular Xtallization (DEMAX) platform of the European Spallation Source ERIC (ESS) moved into Biology House A at Lund University and co-localized with the LP3. DEMAX and LP3 are collaborating to coordinate their efforts to develop cost-effective production of deuterated biomaterials (lipids and proteins) for neutron-based methods such as protein crystallography, neutron reflectometry, and small angle neutron scattering. Perdeuterated proteins will also be crystallized and optimized to promote maximal crystal volumes as required by neutron protein crystallography.
Access to both state-of-the-art large scale facilities (MAX IV for X-rays and ESS for neutrons) will increase the local and international capacity for innovation in the life sciences. Partnering between LP3 and DEMAX enables effective coordination of capabilities in the region. To enable efficient use of these unique and powerful facilities by Lund researchers, Lund University hosts the protein production platform, LP3. LP3 offer services to users from construct design all the way to crystals, if required. The LP3 specializes in protein production, protein purification, biophysical characterization, isotope labelling of both proteins and lipids, and crystallization. DEMAX is an ESS platform dedicated to international users of the neutron scattering facility and will support deuteration of biomolecules and small molecule chemicals, as well as large crystal growth for neutron protein crystallography. The synergies between LP3 and DEMAX are very strong and we look forward to a productive collaboration.
Maria Gourdon, new LP3 scientist responsible for crystallization
Maria Gourdon, PhD, has taken over the responsibility for the crystallization part of LP3. Maria has a background in membrane protein structural biology, handling projects from gene to crystals, structural determination and refinement. Contact her at maria [dot] gourdon [at] biol [dot] lu [dot] se.
New system for microbiological growth monitoring
A Bioscreen C system for microbiological growth studies is now available at LP3. The Bioscreen C can incubate, shake, and measure the turbidity (OD) of up to 200 samples simultaneously.
LP3 and the crystallization facility is "Infrastructure of the month - May"
Protein Production Network Sweden (PPNS) pilot expression screen completed
LP3 is part of the Protein Production Network Sweden (PPNS) that aim to make multiple protein expression hosts, methodologies and competence in protein production available to the Swedish academic life science community. Collaboration between Swedish protein production sites is crucial in order to solve complex scientific questions and in a pilot effort to test a new joint approach, five proteins of varying origin were selected for multi-host expression screening performed the different sites. The screen was competed in November 2014 and a report of the pilot study was made.
A network of Swedish protein production facilities
LP3 is taking part in the formation of a network of Swedish protein production facilities together with the Protein Science Facility at Karolinska Institutet/SciLifeLab, Mammalian Protein Expression at Göteborg University, Protein Expertise Platform at Umeå University and the Drug Discovery and Development platform at SciLifeLab. The network will coordinate optimized use of resources and facilitate increased knowledge transfer.
Eukaryotic expression system
Right now LP3 is in the process of establishing the Baculovirus Expression Vector System (BEVS) – a system for expression of recombinant proteins in insect cells. This system will complement the current LP3 capabilities to express proteins in bacteria and yeast. The BEVS, being a eukaryotic expression system, has its strength in producing proteins from higher eukaryotic organisms that have failed to be produced in E. coli and a strong track record for production of membrane proteins, large proteins and multi-subunit complexes, for structural as well as functional studies.
LP3 is in the process of setting up the Baculovirus Expression Vector System (BEVS).
The LP3 labs are located on floor 1 in Biology building A, Sölvegatan 35, Lund.
Lund Protein Production Platform
Biology Building A
+46 46 2227785
lp3 [at] biol [dot] lu [dot] se
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LP3 acknowledges generous financial support from Lund University and BioCARE (A Strategic Research Area at Lund University).
LP3 and the former crystallization facility acknowledges past generous financial support from Lund University, Knut and Alice Wallenberg Foundation (SWEGENE programme), the Erik & Maja Lundqvist Foundation, the Carl Tesdorpf Foundation and the Swedish Research Council.