Automated Magnetic Separations for Proteomics

Speed and sample purity are major concerns for proteomics researchers, and strategies to achieve higher throughput for sample-handling and data-processing are desired. Magnetic particles comprise a convenient solid support for a variety of assays and procedures based on affinity purification. Paramagnetic beads are especially well-suited for automated procedures because instrumentation exists to easily mix, incubate, and separate the particles in 96-well plates without columns or centrifugation. Such ease of manipulation enables rapid processing of many samples and seamless integration and compatibility with entire workflows.

Figure 1. Highly reproducible IgG purification. Pierce Protein A Magnetic Beads and a KingFisher Instrument were used to purify IgG from rabbit serum in 16 wells of a 96 deep-well plate. Reducing SDS-PAGE results in separate heavy- and light-chain bands of purified IgG. Yield and purity are identical across all wells, confirming the high quality and reproducibility of the beads and instrument.

Figure 2. Consistency of "Pierce High-Capacity Ni-IMAC MagBeads, EDTA Compatible" purifications performed on the KingFisher Flex. Cell culture supernatant (Expi293) containing over-expressed His-tagged HSA (50 mg total protein) was applied to Pierce High-Capacity Ni-IMAC MagBeads, EDTA-Compatible (10ul settled beads) in a 96-well plate. Beads were processed using the KingFisher Flex. Binding was performed with all samples for 30 minutes. The beads were then washed twice, and bound protein was eluted with a 15-minute incubation in Elution Buffer. The eluates were resolved on an SDS-PAGE gel and stained with GelCode Blue (Cat. No. 24594). Gel lanes were normalized to equivalent volume.
M = MW marker, L = lysate load, 1-12 = corresponding elution from plate row. CV’s <10%