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Carbon Dot-Mediated Capillary Electrophoresis Separations of Metallated and Demetallated Forms of Transferrin Protein

1
Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA
2
Razzberry Inc., 5 Science Park, Unit 2E9, New Haven, CT 06511, USA
3
Graduate School of Science and Engineering, Saitama University, Saitama 338-8570, Japan
*
Author to whom correspondence should be addressed.
Molecules 2019, 24(10), 1916; https://doi.org/10.3390/molecules24101916
Received: 2 May 2019 / Revised: 15 May 2019 / Accepted: 16 May 2019 / Published: 18 May 2019
(This article belongs to the Special Issue Analysis of Peptides and Proteins by Electrophoretic Techniques)
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Abstract

Carbon dots (CDs) are fluorescent nanomaterials used extensively in bioimaging, biosensing and biomedicine. This is due in large part to their biocompatibility, photostability, lower toxicity, and lower cost, compared to inorganic quantum dots or organic dyes. However, little is known about the utility of CDs as separation adjuvants in capillary electrophoresis (CE) separations. CDs were synthesized in-house according to a ‘bottom-up’ method from citric acid or other simple carbon precursors. To demonstrate the applicability of CDs as separation adjuvants, mixtures of holo- (metallated) and apo- (demetallated) forms of transferrin (Tf, an iron transport protein) were analyzed. In the absence of CDs, the proteins were not resolved by a simple CE method; however, upon addition of CDs to the separation buffer, multiple forms of Tf were resolved indicating that CDs are valuable tools to facilitate the separation of analytes by CE. CE parameters including sample preparation, buffer identity, ionic strength, pH, capillary inside diameter, and temperature were optimized. The results suggest that dots synthesized from citric acid provide the best resolution of various different forms of Tf and that CDs are versatile and promising tools to improve current electrophoretic separation methods, especially for metalloprotein analysis. View Full-Text
Keywords: carbon dots; capillary electrophoresis; transferrin; metalloproteins; fluorescence carbon dots; capillary electrophoresis; transferrin; metalloproteins; fluorescence
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Sirkisoon, L.R.; Makamba, H.C.; Saito, S.; Colyer, C.L. Carbon Dot-Mediated Capillary Electrophoresis Separations of Metallated and Demetallated Forms of Transferrin Protein. Molecules 2019, 24, 1916.

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