Abstract
AbstractMagnetic microbeads are small iron oxide nanoparticles coated with a bioaffinity material that selectively binds to specific biomolecules of interest, enabling their capture and isolation from complex biological samples. Magnetic microbeads are widely used for purification of specific biomolecules in various experiments in molecular biology. However, current methods of manual pipetting to separate supernatants from magnetic microbeads are often inefficient- time-consuming, labor intensive and inaccurate. Furthermore, the use of pipetting robots and liquid handlers specifically designed for multi-well plates can be a cost-prohibitive approach due to the high cost of equipment and disposable supplies. Here, we developed a centrifugation-based method for high-throughput separation of supernatant from magnetic microbeads. To facilitate the centrifugal separation process, we used the 384 transfer plate™ (Watson, Japan) and a magnetic stand equipped with a 384-well magnetic stand, allowing easy handling of several hundred samples and rapid separation of supernatant from magnetic microbeads. The centrifugal force was used to drive the separation of target molecules from the magnetic microbeads, and sample were successfully separated with relatively high recovery rates. Thus, this technology provides a simple, rapid, and cost- and labor-effective biomolecule separation method with potential applications in various fields, including molecular biology, clinical diagnostics, and biotechnology, and is a valuable addition to the existing toolbox of biomolecule separation methods.
Publisher
Cold Spring Harbor Laboratory
Reference21 articles.
1. Affinity chromatography as a tool for antibody purification;Methods,2012
2. Thirty years of affinity chromatography, React;Funct. Polym,1999
3. Preparation of immobilized chymotrypsin and its use in the isolation of proteolytic inhibitors;Čoupek, Affinity chromatography on hydroxyalkyl methacrylate gels: I;Biochimica et Biophysica Acta (BBA) - Protein Structure,1973
4. Affinity chromatography: A review of trends and developments over the past 50 years, J;Chromatogr. B Analyt. Technol. Biomed. Life Sci,2020
5. characterisation and application of silica-magnetite nanocomposites;J. Magn. Magn. Mater,2004