Abstract
Specific interactions between lectins and glycoproteins determine the outcomes of numerous biological processes. To elucidate the roles of lectins and glycoproteins in those processes, it is essential to detect these proteins in biological samples and purify them to homogeneity. Conventional protein detection and purification techniques are multi-step, time-intensive, and expensive. They often require rigorous trial and error experimentations and fairly larger volumes of crude extracts. To minimize some of these challenges, we recently formulated a new method named Capture and Release (CaRe). This method is rapid, facile, precise, and inexpensive, and it works even when the sample volume is smaller. We developed this method to detect and purify recombinant human Galectin-3 and subsequently validated this method by purifying several other lectins. Besides lectins, CaRe is capable of detecting/purifying glycoproteins. In this method, targets (lectins and glycoproteins) are captured by multivalent ligands called target capturing agents (TCAs). The captured targets are then released and separated from their TCAs to obtain purified targets. CaRe can potentially be used as a tool to discover new lectins and glycoconjugates and elucidate their functions.
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Acknowledgments
This work was supported by National Science Foundation Grant 1608537 (to T.K.D.). A part of this work was supported by a startup fund (to P.B.) and by the Research Excellence Fund (to T.K.D.) provided by Michigan Technological University.
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Kadav, P.D., Edwards, J.L., Krycia, J., Bandyopadhyay, P., Dam, T.K. (2022). Rapid Detection and Purification of Galectin-3 by the Capture and Release (CaRe) Method. In: Stowell, S.R., Arthur, C.M., Cummings, R.D. (eds) Galectins. Methods in Molecular Biology, vol 2442. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2055-7_5
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DOI: https://doi.org/10.1007/978-1-0716-2055-7_5
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