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Design of peptide ligands for affinity purification of human growth hormone
Author(s) -
Chandra Divya,
Timmick Steven,
Goodwine Chaz,
Vecchiarello Nicholas,
Shastry Divya G,
Mullerpatan Akshat,
Trasatti John P,
Cramer Steven,
Karande Pankaj
Publication year - 2019
Publication title -
journal of chemical technology and biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.64
H-Index - 117
eISSN - 1097-4660
pISSN - 0268-2575
DOI - 10.1002/jctb.6029
Subject(s) - chromatography , chemistry , peptide , affinity chromatography , downstream processing , yield (engineering) , ligand (biochemistry) , histidine , adsorption , combinatorial chemistry , biochemistry , materials science , amino acid , organic chemistry , receptor , metallurgy , enzyme
Background In this study, we have demonstrated the design, screening and selection of peptide ligands for the affinity capture of human growth hormone (hGH) from yeast cell cultures. Results Ligand design was carried out using multiple approaches based on primary sequence and structures of natural binding partners of hGH. Screening of potential affinity peptides was conducted using high throughput microarray platforms followed by assessment of in‐solution binding to hGH using fluorescence polarization. Peptide leads were subsequently immobilized on chromatographic resins and the binding and desorption behavior was examined using batch adsorption studies. A lead candidate was examined in further details in column chromatography studies which indicated that while high purity was attained, further refinement was necessary for improved product recovery. Histidine scanning was employed to successfully improve the recovery of hGH from cell culture fluid while still maintaining high purity. Finally, proof‐of‐concept was demonstrated in the column format using complex feed stock where a product purity of 95% was attained at 80% yield. Conclusion The approaches presented here can be translated to other biologics of interest for the rapid development of affinity based purification processes. © 2019 Society of Chemical Industry