Protein purification

The purification of a recombinant protein is the most complex step in the whole service cascade. While cloning of a gene into a transfer vector is often highly dependent on the cDNA fragment size, an efficient purification procedure depends on the biochemical characteristics of the individual protein (e.g. amino acid composition, IP value, glycosylation, stability). In additon individual features of the protein (e.g. secreted, cytoplasmatic or membrane-bound protein, mono- or oligo- or polymeric protein) must be take into account establishing an efficient purification strategy.

Adding a “tag” to a protein (either N- or C- terminally) strongly supports the purification or at least the enrichment of the protein applying standard purification protocols (e.g. using protein A or G Sepharose for the Fc-tag). In addition the broad commercial availability of “tag” - specific antibodies allows an excellent fallback option for protein detection as well as for monitoring all purification steps if antibodies for the specific protein of interest are not available yet. 

 

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With step E of our protein production services in insect cells we support you to design and establish the optimal strategy to purify your protein of interest.

We are looking forward to support you by E-mail or Phone to find a solution suitable for you. Please contact our technical staff for more details.

Note: This part is offered only in combination with step E and a recombinant protein containing a “tag”! For purification of recombinant proteins without an established purification protocol the price will depend on the required time and material needed!

Material we need from you:

Cell pellet or supernatant from service E.

Our service comprises:

  • Preparation of supernatants or cell pellets for purification.
  • Preparation of columns and buffers.
  • Protein purification by affinity chromatography.
  • Analysis of all fractions by SDS-PAGE and/or Western blotting using a „target”- or „tag”-specific antibody.
  • Determination of protein purity by SDS-PAGE and subsequent silver staining.

Results provided to you:

  • Total yield of recombinant protein gained from the volume scale requested by you.
  • A detailed report sheet.

Expected time range:

Dependent on the volume ordered.