TY - JOUR
T1 - Production of functional human insulin-like growth factor binding proteins (IGFBPs) using recombinant expression in HEK293 cells
AU - Wanscher, Anne Sofie Molsted
AU - Williamson, Michael
AU - Ebersole, Tasja Wainani
AU - Streicher, Werner
AU - Wikström, Mats
AU - Cazzamali, Giuseppe
N1 - Copyright © 2014 Elsevier Inc. All rights reserved.
PY - 2015/4
Y1 - 2015/4
N2 - Insulin-like growth factor binding proteins (IGFBPs) display many functions in humans including regulation of the insulin-like growth factor (IGF) signaling pathway. The various roles of human IGFBPs make them attractive protein candidates in drug discovery. Structural and functional knowledge on human proteins with therapeutic relevance is needed to design and process the next generation of protein therapeutics. In order to conduct structural and functional investigations large quantities of recombinant proteins are needed. However, finding a suitable recombinant production system for proteins such as full-length human IGFBPs, still remains a challenge. Here we present a mammalian HEK293 expression method suitable for over-expression of secretory full-length human IGFBP-1 to -7. Protein purification of full-length human IGFBP-1, -2, -3 and -5 was conducted using a two-step chromatography procedure and the final protein yields were between 1 and 12mg protein per liter culture media. The recombinant IGFBPs contained PTMs and exhibited high-affinity interactions with their natural ligands IGF-1 and IGF-2.
AB - Insulin-like growth factor binding proteins (IGFBPs) display many functions in humans including regulation of the insulin-like growth factor (IGF) signaling pathway. The various roles of human IGFBPs make them attractive protein candidates in drug discovery. Structural and functional knowledge on human proteins with therapeutic relevance is needed to design and process the next generation of protein therapeutics. In order to conduct structural and functional investigations large quantities of recombinant proteins are needed. However, finding a suitable recombinant production system for proteins such as full-length human IGFBPs, still remains a challenge. Here we present a mammalian HEK293 expression method suitable for over-expression of secretory full-length human IGFBP-1 to -7. Protein purification of full-length human IGFBP-1, -2, -3 and -5 was conducted using a two-step chromatography procedure and the final protein yields were between 1 and 12mg protein per liter culture media. The recombinant IGFBPs contained PTMs and exhibited high-affinity interactions with their natural ligands IGF-1 and IGF-2.
U2 - 10.1016/j.pep.2014.10.017
DO - 10.1016/j.pep.2014.10.017
M3 - Journal article
C2 - 25448590
SN - 1046-5928
VL - 108
SP - 97
EP - 105
JO - Protein Expression and Purification
JF - Protein Expression and Purification
ER -