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Capturing tagged recombinant proteins

Recombinant proteins are often tagged for purposes of identification and affinity purification, and the same principle can be exploited for capture of recombinant proteins on the sensor surface in Biacore.

Poly-histidine is a commonly used tag that can chelate with Ni²⁺ ions in complex with nitrilotriacetic acid (NTA), providing a convenient approach for capturing his-tagged constructs on Sensor Chip NTA. High affinity antibodies are also available directed against poly-histidine and other commonly used tags, and these can be immobilized on the surface for capture of the tagged proteins.

Another commonly used recombinant tag is glutathione-S-transferase (GST), which may be captured on an anti-GST antibody surface. Antibodies against GST that are suitable for capturing purposes are available from Biacore. Other examples of recombinant tags that may be used for capturing are the FLAG and c-myc epitopes.

The specificity of tag-based capture is valuable in work with recombinant proteins in crude extracts or partially purified samples, since the tag interaction serves the double role of identification and affinity purification of the ligand. This enables characterization of the interaction properties of the recombinant protein directly in crude cell extracts or culture medium, without extensive investment in time and expense to purify the protein.

Lab protocols

Lab procedure Capture of histidine-tagged molecules to Biacore Sensor Chip NTA is available for download on www.cytivalifesciences.com/solutions/protein-research/Interaction-analysis-with-Biacore-surface-plasmon-resonance-SPR/Get-started-with-surface-plasmon-resonance-SPR-interaction-analysis

See also

List of help files

Principles of ligand capture