Prometheus NT.Plex plus NT.Robotic Autosampler

Nanotemper — системы для анализа структуры и взаимодействий биомолекул, купить оборудование Nanotemper в Техноинфо Система анализа аффинности Nanotemper Prometheus NT.Plex plus NT.Robotic Autosampler купить в Техноинфо купить в Техноинфо

Prometheus NT.Plex plus NT.Robotic Autosampler characterizes thermal and chemical unfolding under native conditions using nanoDSF technology. Absolutely label-free, nanoDSF precisely measures the intrinsic fluorescence of a protein while it’s being subjected to either chemical or thermal denaturation.

nanoDSF uses high quality capillaries for a number of reasons — only microliters of sample are required, a wide range of concentrations are quickly measured in-solution, multiple conditions are assessed in one run, and hard-to-measure viscous samples are examined with ease. Just dip a capillary into a sample, place it in Prometheus, and hit start to get Tm, Tonset and Tagg results in minutes. And, if Cm, ΔG and ΔΔG are needed, those can be done too with the same system.

Due to the robust detection method and quick analysis pace, nanoDSF is very versatile and can be used in a number of contexts:

  • Screening of buffers, formulationd, and buffer additives
  • Screening of detergents
  • Analysis of thermal and chemical protein unfolding
  • Long-term protein and antibody storage optimization
  • Forced-degradation stability testing
  • Comparison of biosimilar proteins and antibodies with respect to stability and aggregation
  • Batch-to-batch comparison assays
  • Deep feature analysis (influence of mutations, modifications, conjugations on protein stability and aggregation)

Samples per run: Up to 1536 hands-off

Sample volume measured: 10 μL

Detected molecule concentration range: 0.005 – 250 mg/mL (standard IgG)

Experiment time per run – thermal unfolding (from 1 °С/min up to 7 °С/min ramp): 4 hours to overnight (384 samples)

Experiment time per run – chemical unfolding: 27 minutes (384 samples)

Precision of 1 °С/min thermal ramp: ± 0.2 °С

Temperature control:

  • Ramping options: 0.1 °С/min to 7 °С/min
  • Temperature range: 15 °С – 95 °С (at 25 °С ambient temperature)

Fluorescence detection: 330 nm and 350 nm

Dimensions: 110 cm W x 188 cm H x 90 cm D (stand-alone enclosure)

Weight: 200 kg

Optional upgrades:

  • Aggregation detection optics
  • Plate temperature control

Take assumptions out of the equation — Get reproducible, quantitative results early on from discovery to validation and production. Use Prometheus to generate precise unfolding temperatures (Tm and Tonset), critical denaturant concentrations (Cm), free folding energy (ΔG and ΔΔG), and aggregation onset (Tagg) from start to finish. Be confident about moving forward with the right choice every step of the way.

NanoDSF offers a great number of advantages over traditional fluorimetry approaches. Most importantly, in constrast to standard Differential Scanning Fluorimetry, nanoDSF does not require the used of fluorescent dyes like Sypro Orange.

  • Low sample consumption → Only 10 µL of sample required
  • Free choice of assay buffers → Also biological liquids possible such as serum or cell lysate and other additives/detergents
  • Very short analysis time → enables high throughput
  • Optimal data quality and resolution → Dual 350/330 nm UV-detection
  • Wide temperature range → Analysis possible from 15°C to 95°C
  • No labeling required → Close-to-native analysis possible
  • Wide concentration range → 5 µg/ml to 200 mg/ml
  • Wide molecule size range → From 1 kDa to 1 MDa

NanoDSF is a differential scanning fluorimetry method able to analyze the conformational stability and colloidal stability (aggregation behavior) of proteins under different thermal and chemical conditions. The conformational stability of a protein is described by its unfolding transition midpoint Tm (°C), which is the point where half of the protein is unfolded. The truly label-free nanoDSF technique monitors the intrinsic tryptophan fluorescence of proteins, which is highly sensitive for the close surroundings of the tryptophan residues and which changes upon thermal unfolding.

Up to 64 chips are filled with 10 µl of protein sample and simultaneously scanned at 330/350 nm wavelengths. Melting temperatures are recorded by monitoring changes in the intrinsic tryptophan fluorescence and aggregation onset temperatures are detected via back-reflection light scattering. The samples can be heated to any temperature in the range from 25°C to 95°C. Importantly, samples can be studied without the use of a dye and with free choice of buffer and detergent. Melting temperatures of proteins with a concentration between 5 µg/ml and 250 mg/ml can be analyzed. In order to obtain high quality aggregation onset temperatures, protein solutions with concentrations above 1 mg/ml are required.

Structural basis of α-scorpion toxin action on Nav channels

Thomas Clairfeuille, Alexander Cloake, Daniel T. Infield, José P. Llongueras, Christopher P. Arthur, Zhong Rong Li, Yuwen Jian, Marie-France Martin-Eauclaire, Pierre E. Bougis, Claudio Ciferri, Christopher A. Ahern, Frank Bosmans, David H. Hackos, Alexis Rohou, Jian Payandeh. Science, vol.363, 6433 (2019)

Towards high throughput GPCR crystallography: in meso soaking of adenosine a2a receptor crystals

Prakash Rucktooa, Robert K. Y. Cheng, Elena Segala, Tian Geng, James C. Errey, Giles A. Brown, Robert M. Cooke, Fiona H. Marshall & Andrew S. Doré. Scientific Reports, vol.8, 41 (2018)

High-throughput feasible screening tool for determining enzyme stabilities against organic solvents directly from crude extracts

Severin Wedde, Dr. Christian Kleusch, Dr. Daniel Bakonyi & Prof. Dr. Harald Gröger. ChemBioChem, vol.18(24), p.2399–2403 (2018)

Zinc binding to RNA recognition motif of TDP-43 induces the formation of amyloid-like aggregates

Cyrille Garnier, François Devred, Deborah Byrne, Rémy Puppo, Andrei Yu. Roman, Soazig Malesinski, Andrey V. Golovin, Régine Lebrun, Natalia N. Ninkina & Philipp O. Tsvetkov. Scientific Reports, vol.7, 6812 (2017)

Mutagenesis-independent, stabilization of class B flavin monooxygenases in operation

Goncalves L, Kracher D, Milker S, Rudroff F, Fink M, Ludwig R, Bommarius A, Mihovilovic M. Advanced Synthesis & Catalysis, 359, p.2121–2131 (2017)

Conformational and functional transitions and in silico analysis of a serine protease from conidiobolus brefeldianus (MTCC 5185)

Shukla E, Agrawal S, Gaikwad S. International Journal of Biological Macromolecules, vol.98, (Mtcc 5185) p.387-397 (2017)

Understanding the process-induced formation of minor conformational variants of erwinia chrysanthemi  l-asparaginase

David Gervais, Justin Hayzen, Charlotte Orphanou, Alexandra McEntee, Christine Hallam & Rossalyn Brehm. Enzyme and Microbial Technology, vol.98, p.26-33 (2017)

Creating an efficient methanol-stable biocatalyst by protein & immobilization engineering steps towards efficient biosynthesis of biodiesel

 Shalev Gihaz, Diána Weiser, Adi Dror, Péter Sátorhelyi, Moran Jerabek-Willemsen, Dr. László Poppe, Dr. Ayelet Fishman. ChemSusChem, vol.9, p.3161–3170 (2017)

The deubiquitinase OTULIN is an essential negative regulator of inflammation and autoimmunity

Rune Busk Damgaard, Jennifer A. Walker, Paola Marco-Casanova, Neil V. Morgan, Hannah L. Titheradge, Paul R. Elliott, Duncan McHale, Eamonn R. Maher, Andrew N.J. McKenzie & David Komander. Cell, vol.166, p.1215-1230 (2016)

Structural and functional basis for lipid synergy on the activity of the antibacterial peptide ABC transporter McjD

Mehmood S, Corradi V, Choudhury H, Hussain R, Becker P, Axford D, Zirah S, Rebuffat S, Tieleman D, Robinson C, Beis K. Journal of Biological Chemistry, vol.291 (41), p.21656-21668 (2016)