title: New strategy for the design of ligands for the purification of pharmaceutical proteins by affinity chromatography.
abstract: A new approach for the identification of ligands for the purification of pharmaceutical proteins by affinity chromatography is described. The technique involves four steps. Selection of an appropriate site on the target protein, design of a complementary ligand compatible with the three-dimensional structure of the site, construction of a limited solid-phase combinatorial library of near-neighbour ligands and solution synthesis of the hit ligand, immobilisation, optimisation and application of the adsorbent for the purification of the target protein. This strategy is exemplified by the purification of a recombinant human insulin precursor (MI3) from a crude fermentation broth of Saccharomyces cerevisiae.
authors.affiliation: Institute of Biotechnology, University of Cambridge, UK.
authors.forename: K
authors.initials: K
authors.lastname: Sproule
authors.forename: P
authors.initials: P
authors.lastname: Morrill
authors.forename: J C
authors.initials: JC
authors.lastname: Pearson
authors.forename: S J
authors.initials: SJ
authors.lastname: Burton
authors.forename: K R
authors.initials: KR
authors.lastname: Hejnaes
authors.forename: H
authors.initials: H
authors.lastname: Valore
authors.forename: S
authors.initials: S
authors.lastname: Ludvigsen
authors.forename: C R
authors.initials: CR
authors.lastname: Lowe
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011384:Proinsulin / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae; D014227:Triazines / Q000138:chemical synthesis / Q000737:chemistry
chemical_list: D008024:Ligands; D011994:Recombinant Proteins; D014227:Triazines; D011384:Proinsulin