Journal Highlight: A capillary zone electrophoresis method to detect conformers and dimers of antithrombin in therapeutic preparations

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  • Published: Jul 4, 2016
  • Author: separationsNOW
  • Channels: Proteomics & Genomics
thumbnail image: Journal Highlight: A capillary zone electrophoresis method to detect conformers and dimers of antithrombin in therapeutic preparations
A CZE method, based on a neutral polyethylene oxide-coated capillary and a buffer close to physiological conditions, has been devised for the separation of more than eight forms of antithrombin.

A capillary zone electrophoresis method to detect conformers and dimers of antithrombin in therapeutic preparations

Electrophoresis, 2016, 37, 1696-1703
Anne-Lise Marie, Nguyet Thuy Tran, François Saller, Youmna Mohamed Abdou, Pascal Zeau, Jean-Luc Plantier, Rémi Urbain, Delphine Borgel and Myriam Taverna

Abstract: Antithrombin (AT) is a human plasma glycoprotein that possesses anticoagulant and anti-inflammatory properties. However, the native (active) form of AT is unstable and undergoes conformational changes, leading to latent, cleaved, and heterodimeric forms. The presence of these alternative forms mostly inactive can highly impact the quality and therapeutic activity of pharmaceutical AT preparations. We developed a capillary zone electrophoresis method, based on a neutral polyethylene oxide-coated capillary and a buffer close to physiological conditions, enabling the separation of more than eight forms of AT. Several peaks were identified as native, latent, and heterodimeric forms. The CZE method was reproducible with intraday relative standard deviations less than 0.5 and 2% for migration times and peak areas, respectively. The method was applied to the comparison of AT preparations produced by five competitive pharmaceutical companies, and statistical tests were performed. Important differences in the proportion of each form were highlighted. In particular, one AT preparation was shown to contain a high quantity of heterodimer, and two preparations contained high quantities of latent form. In addition, one AT preparation exhibited additional forms, not yet identified.

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