Skip to Main content Skip to Navigation
Journal articles

Affitins as robust tailored reagents for affinity chromatography purification of antibodies and non-immunoglobulin proteins

Abstract : Affinity chromatography is a convenient way of purifying proteins, as a high degree of purity can be reached in one step. The use of tags has greatly contributed to the popularity of this technique. However, the addition of tags may not be desirable or possible for the production of biopharmaceuticals. There is thus a need for tailored artificial affinity ligands. We have developed the use of archaeal extremophilic proteins as scaffolds to generate affinity proteins (Affitins). Here, we explored the potential of Affitins as ligand to design affinity columns. Affitins specific for human immunoglobulin G (hIgG), bacterial PulD protein, and chicken egg lysozyme were immobilized on an agarose matrix. The columns obtained were functional and highly selective for their cognate target, even in the presence of exogenous proteins as found in cell culture media, ascites and bacterial lysates, which result in a high degree of purity (∼95%) and recovery (∼100%) in a single step. Anti-hIgG Affitin columns withstand repetitive cycles of purification and cleaning-in-place treatments with 0.25 M NaOH as well as Protein A does. High levels of Affitin productions in Escherichia coli makes it possible to produce these affinity columns at low cost. Our results validate Affitins as a new class of tailored ligands for the affinity chromatography purification of potentially any proteins of interest including biopharmaceuticals.
Document type :
Journal articles
Complete list of metadatas

Cited literature [36 references]  Display  Hide  Download

https://www.hal.inserm.fr/inserm-01292506
Contributor : Elizabeth Bernardo <>
Submitted on : Wednesday, March 23, 2016 - 11:57:17 AM
Last modification on : Friday, June 19, 2020 - 3:08:40 AM
Long-term archiving on: : Monday, November 14, 2016 - 2:33:50 AM

File

 Restricted access
To satisfy the distribution rights of the publisher, the document is embargoed until : jamais

Please log in to resquest access to the document

Identifiers

Collections

Citation

Ghislaine Béhar, Axelle Renodon-Cornière, Barbara Mouratou, Frédéric Pecorari. Affitins as robust tailored reagents for affinity chromatography purification of antibodies and non-immunoglobulin proteins. Journal of Chromatography A, Elsevier, 2016, 1441, pp.44-51. ⟨10.1016/j.chroma.2016.02.068⟩. ⟨inserm-01292506⟩

Share

Metrics

Record views

183