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Recombinant protein purification from pea
Author(s) -
Menkhaus Todd J.,
Pate Cynthia,
Krech Anthony,
Glatz Charles E.
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20039
Subject(s) - isoelectric point , recombinant dna , chromatography , chemistry , affinity chromatography , isoelectric focusing , histidine , downstream processing , protein precipitation , protein purification , precipitation , ion chromatography , solubility , biochemistry , high performance liquid chromatography , amino acid , organic chemistry , enzyme , physics , meteorology , gene
To assess the suitability of transgenic peas as a host for protein production from the perspective of ease of recovery, a strain containing recombinant β‐glucuronidase with poly(histidine) tail (GUSH6) was evaluated for solubility of the target protein in relation to native components (proteins, carbohydrates, and phenolics). Recovery of the recombinant GUSH6 from aqueous extracts by immobilized metal affinity chromatography with coupled Co 2+ yielded a nearly pure product with IDA (enrichment factor (EF) = 260) or NTA (EF = 200) resin. Single‐step recoveries were also possible by isoelectric precipitation (EF = 4), polyelectrolyte precipitation (EF = 1.5), and anion‐exchange chromatography (EF = 3.1), but enrichment factors were low. © 2004 Wiley Periodicals, Inc.