LP3 -

Lund Protein Production Platform

Infrastructure

Research areas and keywords

UKÄ subject classification

  • Medical and Health Sciences
  • Engineering and Technology
  • Natural Sciences
  • Agricultural Sciences
  • Agricultural Biotechnology
  • Other Chemical Engineering
  • Pharmaceutical Chemistry
  • Bio Materials
  • Biocatalysis and Enzyme Technology
  • Bioprocess Technology
  • Medical Biotechnology
  • Other Industrial Biotechnology
  • Pharmaceutical Biotechnology
  • Cell and Molecular Biology
  • Medicinal Chemistry
  • Other Basic Medicine
  • Other Medical Biotechnology
  • Biomaterials Science
  • Biological Sciences
  • Chemical Sciences
  • Other Natural Sciences
  • Biochemistry and Molecular Biology
  • Biophysics
  • Structural Biology
  • Other Chemistry Topics

Type of infrastructure

  • Equipment
  • Services

Degree of recognition

  • Local
  • National
  • International

Keywords

  • Protein Production, Structural Biology, Protein Structure, Stable isotope labeling, Deuteration, Biophysics, Baculovirus vector expression system ( BEVS ), Fragment screening, Crystallography, Protein Purification, Protein Expression, Neutron, Neutron scattering, DSL, DSF

Name of national/international infrastructure this infrastructure belongs to

Lund Protein Production Platform (LP3) is a cross-faculty center of Lund University for protein production, crystallization and structure determination (www.lu.se/lp3). Its staff covers the whole process chain from protein production to solving and refining protein structures. The platform is currently headed and managed by Dr. W. Knecht.

LP3 participates in relevant national and international networks and societies (PPNS (http://www.ppns.ki.se), ARBRE-MOBIEU (https://arbre-mobieu.eu), P4EU (https://p4eu.org) and CTLS (http://www.ctls-org.eu/)).

The DEuteration and MAcromolecular Xtallization (DEMAX) platform of the European Spallation Source ERIC (ESS) is 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.

LP3 is part of the FragMAX Project (“FragMAX - a facility for high throughput fragment screening in drug development by X-ray crystallography”) aiming to set up fragment screening at the BioMAX beamline of the MAX IV laboratory.

Description

LP3’s mission is to:
- offer open service and support, primarily to researchers at LU, with protein production, characterization and crystallization for their research projects.
- be responsible for a common and open infrastructure for protein production and crystallization, as well as to contribute actively to the interaction of LU with MAX IV, ESS and other relevant major research facilities, networks and initiatives.
- if needed, to act as LU’s node in a national infrastructure in the protein science area.
- develop competence and methods in the area of protein sciences.
- serve the surrounding community (e.g. closely located large infrastructures, small biotech etc.)

Equipment and resources

Lund Protein Production Platform (LP3) is a cross-faculty center of Lund University for protein production, crystallization and structure determination (www.lu.se/lp3). The center is run by a manager and 6 research engineers. 3 senior lecturers are associated with LP3. The staff covers the whole process chain from protein production to solving and refining protein structures. The platform is currently headed and managed by Dr. W. Knecht.
The center is fully equipped for recombinant protein production in E. coli and insect cells (BEVS) and protein purification using state of the art chromatography systems. Equipment for SDS-PAGE, Western blotting and other standard equipment for protein characterization, biophysical characterization as DSF and DSL and enzymatic activity assays is available at the center or within close proximity. All documentation is captured using electronic lab notebooks.
For crystallization, the facility is equipped with state-of-the-art nanolitre pipetting equipment with the recently-added capability to handle lipidic cubic phases for membrane proteins, as well as two “plate hotels” with the capacity to store and automatically image up to 600 or 256 plates, each with up to 288 crystallization trials, at 4 °C and 20 °C, respectively. A Tecan liquid handling system and a Dragonfly liquid handler for the preparation of focused crystallization screens are also available. For optimization of seeding conditions we have access to an Oryx8.
LP3 is colocalized with and closely collaborates with the DEMAX platform of the ESS on production of deuterium labeled biomolecules (proteins, lipids) for neutron scattering experiments and the development of methods for crystal growth for neutron crystallography. LP3 is to our knowledge currently the only general academic protein production platform in Sweden providing specialization in protein production in the BEVS and perdeuteration of biological macromolecules.
Moreover, LP3 has currently regularly access to beamtime at the MAX IV laboratory BioMAX beamline for testing crystals produced at LP3 and to collect datasets. LP3 has also all competence to further process data, solve and refine protein structures.
This makes LP3 to an unique entry point and partner for both research aiming at conventional x-ray protein structure determination, but also for enabling research using neutron scattering techniques that require deuterated bioreagents.

Services provided

LP3 offers services for the entire process chain of production, purification, characterization, protein crystallization and protein structure determination and structure refinement or each individual step in the chain. LP3 can help with:

•Plasmids for protein production

•Microbiological growth monitoring (Bioscreen C)

•Recombinant protein production in bacterial (E. coli) or eukaryotic (insect) cells.

•Protein labeling (seleno-methionine incorporation, labeling with stable isotopes (2H, 13C, 15N), biotinylation, phosphorylation)

•Protein purification

•Biophysical protein characterization by Size Exclusion Chromatography (SEC), Dynamic Light Scattering (DLS) and Differential Scanning Fluorimetry (DSF)

•High-throughput & nanovolume protein crystallization

•Automated crystallization plate storage and imaging at 4 and 20 °C

Protein structure determination and refinement:

•Application for beamtime, however LP3 is part of a BAG for BioMAX.

•MX data collection at synchrotron beamline (BioMAX MAX IV)

•Process x-ray data and determine and refine the structures

•Cryo-EM screening, LP3 is part of a BAG for Cryo-EM in Stockholm

For details of current services, please see LP3 homepage: www.lu.se/lp3

Management of the infrastructure

LP3 is governed by a board of one chairman and 6 members, one each from Faculty of Science (FoS), Faculty of Medicine (FoM), LTH, MAX IV and ESS (external member) and one student. The chairman is the dean or vice dean of the FoS. The daily business of the center is led by a manager (Dr. Wolfgang Knecht).

LP3 is placed at the Biology Department (Biology Building A, Sölvegatan 35, 22362 Lund), within the FoS at LU. LP3 is a separate entity within the existing administrative structure of the Department of Biology and follows the working and delegation principles of the FoS.

Available for loan

Available for loan - internal and external

Terms of access:

LP3 offers open service and support, primarily to researchers at LU, with protein production, protein characterization, protein crystallization and protein structure determination for their research projects. However external users can also approach LP3 (see mission statement).<br/><br/><b>For protein production (with or without crystallization):</b><br/>An e-mail can be send to lp3@biol.lu.se to initiate a discussion about the project in question. In view of the customized nature of the services provided it is necessary to obtain a quotation from LP3 for the desired work.<br/><br/><b>For crystallization only:</b><br/>A price list for protein crystallization and characterization can be found here: http://www.biology.lu.se/services/lp3-lund-protein-production-platform/protein-crystallization-and-characterization<br/><br/>

Recent research outputs

Cassimjee, K. E., Hendil-Forssell, P., Volkov, A., Krog, A., Malmo, J., Aune, T. E. V., Wolfgang Knecht, Miskelly, I. R., Moody, T. S. & Svedendahl Humble, M., 2017 Dec 6, In : ACS Omega. 2, 12, p. 8674-8677

Research output: Contribution to journalArticle

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