Adsorption and immobilization of cytochrome c on nanodiamonds.
Methods have been developed to immobilize proteins onto the surfaces ofnanodiamonds with an average size of 5 +/- 1 nm. The immobilization started with carboxylation/oxidization of diamonds with strong acids, followed by coating the surfaces with poly-L-lysine (PL) for covalent attachment of proteins using heterobifunctional cross-linkers. The feasibility of this approach is proven with fluorescent labeling of the PL-coated diamonds by Alexa Fluor 488 and subsequent detection of the emission using a confocal fluorescence microscope. Immobilization of proteins onto the surfaces is also demonstrated with yeast cytochrome c, which possesses a free SH group for linkage and a characteristic Soret absorption band for observation.
Huang
L C Lora
LC
Institute of Atomic and Molecular Sciences, Academia Sinica, P.O. Box 23-166, Taipei, Taiwan 106, Republic of China.
Chang
Huan-Cheng
HC
D000327:Adsorption; D045304:Cytochromes c / Q000737:chemistry*; D018130:Diamond / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005240:Feasibility Studies; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D049329:Nanostructures / Q000737:chemistry*; D010084:Oxidation-Reduction; D010316:Particle Size; D011107:Polylysine / Q000138:chemical synthesis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013499:Surface Properties
D004800:Enzymes, Immobilized; D011107:Polylysine; D018130:Diamond; D045304:Cytochromes c