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

Molecules

LeonaR. Sirkisoon¹  ChristaL. Colyer¹  Shingo Saito²  HonestC. Makamba³ 

[1] Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA;Graduate School of Science and Engineering, Saitama University, Saitama 338-8570, Japan;Razzberry Inc., 5 Science Park, Unit 2E9, New Haven, CT 06511, USA;

关键词: carbon dots;    capillary electrophoresis;    transferrin;    metalloproteins;    fluorescence;   

DOI  :  10.3390/molecules24101916

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown