Product
제품
- Calypso®
- 제품명: Calypso®
- 용도: CG-MALS
- 메이커: Wyatt Technology
- 카달로그:
소개
Calypso®
Label-Free, Immobilization-Free Characterization Of Protein-Protein and
Other Macromolecular Interactions with Composition-Gradient Multi-Angle Light Scattering.
The Calypso® II, in conjunction with a DAWN® or miniDAWN® MALS detector, measures binding affinities and absolute, molecular stoichiometries of complex biomolecular interactions. In addition to equilibrium association constants Kd and absolute stoichiometry of reversible self- and hetero-associations, it can also help determine reaction or aggregation rates, non-specific interaction parameters (virial coefficients) and even automate measurements of weight-average molar mass and dn/dc for polymers. Included with each Calypso is a copy of CALYPSO software, the most versatile software package available for analysis of biomolecular interactions in solution by light scattering.
Creating composition gradients...
CG-MALS employs a Calypso connected to a MALS instrument and optional concentration detector. The Calypsoperforms sample preparation and delivery, combining up to three different solution in precise mixing ratios and injecting into the detectors. Accurate compositions result from setting the Calypso's pumps to appropriate flow rate ratios, allcontrolled from the CALYPSO™ software through an intuitive GUI. Mixing is achieved by dispensing the solutions simultaneously from all three pumps via a static mixer.
...that work with MALS detectors
Reliable light scattering measurement require the samples to be degassed and filtered. The Calypsoincorporates built-in, low-volume degassers and filter housings, connected via PEEK tubing andfinger-tight fittings. Even the filter housing is finger-tight, making use of a proprietary design for maximum convenience.
Calypso features
• Biocompatible wetted materials.
• Flexibility to re-organize fluid paths and utilize different wetted materials as needed.
• Convenient loading of sample and diluent solutions via conical tubes: 5, 15 or 50 mL.
• Autoinject Out contact closure signal, to trigger data acquisition by ASTRA® or DYNAMICS® software fordata acquisition and analyses not supported by
the CALYPSO software (e.g., DLS acquisition).
• Built-in Wash port, allowing the pumps to draw on reservoirs of wash solutions for post-experimental clean-up. Automation of up to two sequential wash
solutions, e.g. buffer (to clean out proteins) followed by water (to clean out buffer salts) or detergent followed by water, is possible when an
Orbit solvent selector valve is added to the setup. These operations are appended to the Method for unattended operation.
About CG-MALS
Composition-Gradient Multi-Angle Light Scattering (CG-MALS) employs a series of unfractionated samples of different composition or concentration in order to characterize a wide range of macromolecular interactions. No special modifications (e.g., sample tagging or immobilization procedures) are necessary: samples are unlabeled and entirely in solution.
Addressing a host of biomolecular phenomena
The primary analysis techniques supported by Calypso hardware and software are:
• Dynamic equilibria: specific binding and complex assemblies of proteins, oligonucleotides and other biomolecules; KD (equilibrium dissociation constant)
from picomolars to millimolars; absolute molecular stoichiometry of associating complexes; self and/or heteroassociations
• Non-specific macromolecular interactions: self- and cross-virial coefficients
• Stop flow kinetics: aggregation, dissociation and other time-dependent reactions: equilibration time from seconds to hours
• Zimm plots: concentration gradients for determining solution-average molar mass MW, size Rg, and second virial coefficients
• Refractive index increment:dn/dc
Calypso's automation enhances productivity while the CALYPSO software provides an unparalleled selection of interaction models. Relative to other techniques for characterizing protein interactions, such as surface plasmon resonance, sedimentation equilibrium, kinetic exclusion or isothermal titration calorimetry – as well as manual CG-MALS measurements – the Calypso provides fast and accurate results.
Applications
Drug Discovery
• Quantify binding affinity and stoichiometry of enzyme/inhibitor or antibody/antigen interactions, including complex multi-valent and multi-protein
complex formation
• Study the impact of small molecules on protein-protein interactions
Process Improvement
• Determine second virial coefficient and adjust buffer parameters to improve formulation stability and viscosity
• Determine cross virial coefficients to optimize antibody purification and understand the effects of large excipients on formulations
Self-assembly / Aggregation
• Quantify impact of solvent ionic strength, pH, or excipients on polymerization or protein associations
• Measure kinetics of self-assembly and aggregation via rate of change of molar mass and radius of gyration, and hydrodynamic radius
(with a WyattQELS module or NanoStar and parallel analysis in ASTRA or DYNAMICS)
Biotech R&D
• Characterize macromolecular binding affinity and associated complex stoichiometry over a wide range of buffer compositions, time, and temperature scales
사양
액세서리
Orbit - The Orbit helps conserve mobile phase by programmatically directing the solvent to either a waste bottle or back to the solvent reservoir. In conjunction with the Calypso, the Orbit is used to programmatically choose from one of two Wash solutions in order to enhance automated post-experimental clean-up.
Solaris - Prevents growth of bacteria, fungi or algae in solvents with high efficiency and protects your system from down-time due to contamination.
관련자료
Protein-Ligand Binding
Most protein-ligand and similar biomolecular interactions are best measured at or below 1 mg/mL. The CG-MALS configuration should include:
- Calypso
- DAWN H/C for temperature regulation of the interaction and highest sensitivity, though a miniDAWN (ambient only) may suffice
- Third-party on-line UV/Vis absorption detector - please Contact Support for recommended models
Non-Specific Interactions
Non-specific interactions are usually measured in the range of 1 - 25 mg/mL. The configuration should include:
- Calypso
- DAWN H/C when measuring temperature-sensitive samples or polymers with Rg > 40 nm; otherwise miniDAWN
- Optilab for concentration measurements
High-Concentration Protein-Protein Interactions
These measurements may go to concentrations of hundreds of mg/mL protein.
- Calypso
- DAWN H/C when measuring temperature-sensitive samples or polymers/aggregates with Rg > 40 nm; otherwise miniDAWN
- Optilab HC (high-concentration option) for concentration measurements
- Batch Conversion Kit and microCUVETTE for concentrated, viscous solutions that cannot be run through the Calypso and MALS flow cell