New strategy for the design of ligands for the purification of pharmaceutical proteins by affinity chromatography

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 740; no. 1; pp. 17 - 33
• Main Authors: Sproule, Kenny, Morrill, Paul, Pearson, James C, Burton, Steven J, Hejnæs, Kim R, Valore, Henrik, Ludvigsen, Svend, Lowe, Christopher R
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 31.03.2000; Elsevier Science
• Subjects: Amino Acid Sequence; Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Binding Sites; Biological and medical sciences; Chromatography, Affinity - methods; Fundamental and applied biological sciences. Psychology; Humans; Insulin; Ligands; Models, Molecular; Molecular Sequence Data; Proinsulin - genetics; Proinsulin - isolation & purification; Proteins; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Saccharomyces cerevisiae; Triazines - chemical synthesis; Triazines - chemistry; Proteins; Ligands; Insulin; Chemical combinatorial library; Separation method; Purification; Three dimensional structure; Affinity chromatography; Ligand; Combinatorial chemistry; Recombinant protein
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(99)00570-8

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.