Water-soluble quantum dots (QDs) were used to label goat anti-human immunoglobulin antibodies (Abs), and the labeling process was characterized by column purification. The QDs obtained in organic solvent were modified with mercaptoacetic acid (MAA) and became water-soluble. These water-soluble QDs were linked to the antibodies using the coupling reagents ethyl-3-(dimethyl aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The linking process was shown to be effective by ultra-filter centrifugation and column purification. After comparing the quantities of Abs and water-soluble QDs involved in the linking reaction via column purification, it was found that a molar Abs:QD ratio of >1.2 resulted in most of the water-soluble QDs becoming covalently linked to the Abs. The circular dichroism (CD) spectra of Abs and QD-Ab conjugates were very similar to each other, indicating that the secondary structure of Abs remained largely intact after the conjugation. Finally, antigen (Ag)-antibody (Ab) recognition reactions perfomed on the surface of a glass slide showed that the conjugate retained the activity of Abs. This work lends support to the idea of linking biomolecules to QDs, and thus should aid the application of QDs to the life sciences.
CdTe quantum dots (QDs) were synthesized in aqueous solution with 3-mercaptopropionic acid as the stabilizer. Chemically reduced bovine serum albumin (BSA) was used to modify the surface of the QDs. Experimental results showed that the denatured BSA (dBSA) could be effectively conjugated to the surface of CdTe QDs. Column chromatography was used to purify the conjugates and determine the optimal ratio of dBSA to QDs. Further experimental results showed that the conjugation of QDs by dBSA efficiently improved the photoluminescence quantum yield, the chemical stability of QDs and their stability against photobleaching. A facile and sensitive method for determination of silver(I) ions was proposed based on the fluorescence quenching of the dBSA-QDs. Under the optimal conditions, the relative fluorescence intensity decreased linearly with the concentration of the silver(I) ions in the range 0.08-10.66 microM. The detection limit was 0.01 microM. This study provides a new method for the detection of metal cations.
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