title: Automated operation of immiscible filtration assisted by surface tension (IFAST) arrays for streamlined analyte isolation.
abstract: The purification of analytes is an important prerequisite for many analytical processes. Although automated infrastructure has dramatically increased throughput for many of these processes, the upstream analyte purification throughput has lagged behind, partially due to the complexity of conventional isolation processes. Here, we demonstrate automated operation of arrays of a new sample preparation technology--immiscible filtration assisted by surface tension (IFAST). IFAST uses surface tension to position an immiscible liquid barrier between a biological sample and downstream buffer. Paramagnetic particles are used to capture analytes of interest and draw them across the immiscible barrier, thus resulting in purification in a single step. Furthermore, the planarity of the IFAST design enables facile and simultaneous operation of multiple IFAST devices. To demonstrate the application of automation to IFAST, we successfully perform an array of 48 IFAST-based assays to detect the presence of a specific antibody. This assay array uses only a commercial automated liquid handler to load the devices and a custom-built magnet actuator to operate the assays. Automated operation of the IFAST devices resulted in more repeatable results relative to manual operation.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA. scott.michael.berry@gmail.com
authors.forename: Scott M
authors.initials: SM
authors.lastname: Berry
authors.forename: Keil J
authors.initials: KJ
authors.lastname: Regehr
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Casavant
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000302:isolation & purification; D057205:Automation, Laboratory; D002623:Chemistry Techniques, Analytical; D045424:Complex Mixtures / Q000737:chemistry*; D005374:Filtration / Q000295:instrumentation* / Q000379:methods; D057166:High-Throughput Screening Assays; D006801:Humans; D060328:Magnetic Phenomena; D046228:Microarray Analysis / Q000706:statistics & numerical data*; D013500:Surface Tension
chemical_list: D000906:Antibodies; D045424:Complex Mixtures