Characterization and application of quantum dot nanocrystal—monoclonal antibody conjugates for the determination of sulfamethazine in milk by fluoroimmunoassay

Jianzhong Shen; Fei Xu; Haiyang Jiang; Zhanhui Wang; Jing Tong; Pengju Guo; Shuangyang Ding
December 2007
Analytical & Bioanalytical Chemistry;Dec2007, Vol. 389 Issue 7/8, p2243
Academic Journal
Quantum dot (Qdot) nanocrystals have been increasingly used as fluorescence labels in fluoroimmunoassays recently because of their excellent optical characteristics. In this paper, a new monoclonal antibody (MAb) against sulfamethazine (SMZ) was successfully produced and linked to Qdot nanocrystals by covalent coupling. The Qdot–MAb conjugates were characterized by SDS-PAGE and high-performance capillary electrophoresis (HPCE). An enzyme-linked immunosorbent assay (ELISA) method was utilized to evaluate the antigen–antibody binding affinity and then a novel direct competitive fluorescence-linked immunosorbent assay (cFLISA) for the detection of SMZ in milk by using Qdots as fluorescent labels was evaluated. The results showed that the 50% inhibition values (IC50) of the cFLISA were 4.3 ng/mL in milk and 5.2 ng/mL in PBS, and the limits of detection (LODs) were 0.6 ng/mL in milk and 0.4 ng/mL in PBS, respectively. The recoveries of SMZ from spiked milk samples at levels of 10–100 ng/mL ranged from 94 to 106%, with coefficients of variation (CVs) of 2.1–9.2%. [Figure not available: see fulltext.]


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