Recombinant Protein Purification

Recombinant Protein Purification

The Facility provides analytical and preparative scale, one- or two-step purification of recombinant proteins expressed in bacteria and baculovirus infected insect cells. The expectation is to provide highly purified recombinant proteins for use in structure-function studies, crystallographic efforts, assay development for high-throughput small molecule screening, custom antibody productions, and peptide identification of macromolecular protein complexes. Since most recombinant protein expression in bacterial and baculovirus systems takes advantage of epitope tags for monitoring expression, proteins are purified from soluble extracts using appropriate affinity matrices (e.g., Ni2+-/Co2+-agarose, GSH-sepharose, amylose-agarose, anti-FLAG/HA agarose) that allow for selection of specific tags (e.g. 6His, GST).

When necessary, a second purification step (e.g., affinity, size exclusion chromatography, etc) is included to ensure maximum purity of the recombinant protein. The core cleaves epitope tags, when requested, and the recombinant protein will be further purified from tags and proteases. Purified proteins are dialyzed into appropriate buffers and concentrated to desired concentrations.

Timeline:

~1-2 weeks

Required Materials:

Cellular source of recombinant protein expression

Deliverables:

  • Purified protein dialyzed into desired storage buffer
  • Experimental reports and QC data