Zimm Plot/Second Virial Coefficient (A2)
Light scattering can be used to measure the second virial coefficient (A2) of a macromolecule. A2 is a measure of macromolecular self association, and is one of the few parameters that can be used to predict the crystallization properties of a sample. A2 is also an important parameter for characterizing protein-protein interactions. Moreover, light scattering techniques used to measure A2 can be extended to characterize hetero-association between two different proteins.
Traditionally, A2 was measured using a painstaking process of preparing multiple concentrations of a sample, and then making multiple measurements to construct a classic Zimm plot. Wyatt Technology is committed to bringing this technology into the 21st century, particularly since it is of growing importance to the protein characterization community. Some of the avenues that we have implemented or are exploring include:
Online A2 determination
ASTRA supports the analysis of rapid injections of small volumes of a sample, ideally prepared in an autodilution process in an autosampler. This proprietary online analysis can retrieve A2 using a fraction of the sample needed for traditional measurements, and supports automation and high-throughput studies.
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Global fitting Traditional Zimm plots involved multiple fits to multiple sets of data, and extrapolations to zero scattering angle and concentration. ASTRA employs a proprietary global fitting technology that upgrades the Zimm plot to a more robust, modern analysis. The global fitting technology removes all extrapolations from the Zimm plot method, helps identify inconsistent data sets, and provides more precise and robust results for A2. As with all analysis procedures in ASTRA, the global fit view (shown at right) offers immediate visual confirmation of fit quality using a new visual representation of the Zimm plot. Parameters can be changed in the grid view, and fit results viewed immediately. |
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Hetero-association studies
Wyatt Technology is very excited about the prospects of measuring hetero-association between different proteins or macromolecules by extending the A2 analysis. Please contact Wyatt Technology for more information if you are interested in hetero-association studies.
