Competitive immunoassay for recombinant hirudin using capillary electrophoresis with laser-induced fluorescence detection.
作者： Eunmi Ban , Hui-Sun Nam , Young Sook Yoo
DOI: 10.1016/S0021-9673(01)00829-9
关键词:
摘要: A competitive immunoassay based on capillary electrophoresis (CE) with laser-induced fluorescence (LIF) has been developed for the determination of recombinant hirudin (r-hirudin) in biological mixtures. Hirudin, a thrombin inhibitor, is polypeptide 65 amino acids. To check purity levels and perform pharmacokinetic studies (r-hirudin), specific reproducible analysis methods are demanded. The work involved development separation conditions allowing routine plasma samples. In this study, r-hirudin was labeled fluorescein isothiocyanate (FITC), FITC-labeled purified using high-performance liquid chromatography. product then mixed sample followed addition anti-hirudin antibody. Free, antibody-bound, tagged could be separated within 5 min by CE uncoated fused-silica high reproducibility. method can used to determine good precision detection limit lower than 20 nM. This result demonstrates feasibility CE-LIF
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