Ban, E Nam, HS Yoo, YS 2024-01-21T12:05:50Z 2024-01-21T12:05:50Z 2022-01-10 2001-07-27 0021-9673 https://pubs.kist.re.kr/handle/201004/140305 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 a polypeptide of 65 amino acids. To check purity levels and perform pharmacokinetic studies of (r-hirudin), specific and reproducible analysis methods are demanded. The work involved the development of separation conditions allowing for routine analysis of plasma samples. In this study, r-hirudin was labeled with fluorescein isothiocyanate (FITC), and FITC-labeled r-hirudin was purified using high-performance liquid chromatography. The purified product was then mixed with the sample followed with the addition of anti-hirudin antibody. Free, antibody-bound, and tagged r-hirudin could be separated within 5 min by CE analysis using uncoated fused-silica capillary with high reproducibility. The developed method can be used to determine r-hirudin with good precision and a detection limit lower than 20 nM. This result demonstrates the feasibility of the CE-LIF immunoassay method for the determination of r-hirudin in plasma samples. (C) 2001 Elsevier Science B.V. All rights reserved. English ELSEVIER SCIENCE BV DERIVATIVES MANILLENSIS ASSAY ELISA Competitive immunoassay for recombinant hirudin using capillary electrophoresis with laser-induced fluorescence detection Article 10.1016/S0021-9673(01)00829-9 1 JOURNAL OF CHROMATOGRAPHY A, v.924, no.1-2, pp.337 - 344 JOURNAL OF CHROMATOGRAPHY A 924 1-2 337 344 scie scopus 000170359200034 2-s2.0-0035958537 Biochemical Research Methods Chemistry, Analytical Biochemistry & Molecular Biology Chemistry Article; Proceedings Paper DERIVATIVES MANILLENSIS ASSAY ELISA immunoassays hirudin polypeptides