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Automated High-Throughput Protein Purification with MagneHis™
Particles
T.V.S. Murthy, Zhenwei Shi, Mauricio Fernandez and
Leonardo Brizuela
Institute of Proteomics, Harvard Medical School
High-throughput protein production facilitates the study of multiple
families of proteins involved in various metabolic pathways. Purified
proteins find innumerable applications in biochemical and immunological
assays for therapeutic discovery and diagnostics. In order to generate
purified proteins for these applications, we developed an automated
high-throughput protein purification pipeline. The basic elements of
this 96-well format pipeline include generating clones and expressing
and purifying proteins. Cell growth and expression are performed in
96-well, deep-well blocks using liquid handling robots where necessary.
We have completely automated the protein purification process on the
Biomek® FX using a magnetic bead-based affinity purification
procedure. In this report, we briefly describe the process of
robotic-affinity protein purification and present our purification
results with forty-eight polyhistidine-tagged proteins. The pipeline
consistently yielded greater than 10µg of protein with ~90% purity from
1ml cultures.
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