Expression and purification of biologically active IGF-binding proteins using the LCR/Mel expression system

Wendy Bagnall; Paul M. Sharpe; Peter Newham; Johnathan Tart; Richard A. Mott; Vanessa R. Torr; Robert A. Forder; M. R C Needham

doi:10.1016/S1046-5928(02)00549-1

Expression and purification of biologically active IGF-binding proteins using the LCR/Mel expression system

Wendy Bagnall, Paul M. Sharpe, Peter Newham, Johnathan Tart, Richard A. Mott, Vanessa R. Torr, Robert A. Forder, M. R C Needham

Research output: Contribution to journal › Article › peer-review

Abstract

The anabolic effects and bioavailability of insulin-like growth factors I and II (IGF-I, IGF-II) are regulated in part by a family of IGF-binding proteins (IGFBPs). There are six known members of the IGFBP family, which share distinct structural characteristics and functional activities. To study the binding properties of these proteins, we have expressed recombinant IGFBP-3 and IGFBP-4 using the LCR/Mel expression system. Using this system, we found that recombinant IGFBP-3 was secreted by Mel cells and had a glycosylation pattern similar to that of native IGFBP-3. Recombinant IGFBP-4 secreted from Mel cells had a molecular size identical to that of non-glycosylated native IGFBP-4. The binding kinetics of recombinant IGFBPs was measured using a solid-phase ligand-binding assay, an in vitro solution-binding assay, and a cellular proliferation assay. IGF-I bound with high affinity to recombinant IGFBP-3 and IGFBP-4 with KDs of

Original language	English
Pages (from-to)	1-11
Number of pages	10
Journal	Protein Expression and Purification
Volume	27
Issue number	1
DOIs	https://doi.org/10.1016/S1046-5928(02)00549-1
Publication status	Published - 1 Jan 2003

Access to Document

10.1016/S1046-5928(02)00549-1

Cite this

@article{fd9935acc2264413a5c19e92bf42fe9c,

title = "Expression and purification of biologically active IGF-binding proteins using the LCR/Mel expression system",

abstract = "The anabolic effects and bioavailability of insulin-like growth factors I and II (IGF-I, IGF-II) are regulated in part by a family of IGF-binding proteins (IGFBPs). There are six known members of the IGFBP family, which share distinct structural characteristics and functional activities. To study the binding properties of these proteins, we have expressed recombinant IGFBP-3 and IGFBP-4 using the LCR/Mel expression system. Using this system, we found that recombinant IGFBP-3 was secreted by Mel cells and had a glycosylation pattern similar to that of native IGFBP-3. Recombinant IGFBP-4 secreted from Mel cells had a molecular size identical to that of non-glycosylated native IGFBP-4. The binding kinetics of recombinant IGFBPs was measured using a solid-phase ligand-binding assay, an in vitro solution-binding assay, and a cellular proliferation assay. IGF-I bound with high affinity to recombinant IGFBP-3 and IGFBP-4 with KDs of",

author = "Wendy Bagnall and Sharpe, {Paul M.} and Peter Newham and Johnathan Tart and Mott, {Richard A.} and Torr, {Vanessa R.} and Forder, {Robert A.} and Needham, {M. R C}",

year = "2003",

month = jan,

day = "1",

doi = "10.1016/S1046-5928(02)00549-1",

language = "English",

volume = "27",

pages = "1--11",

journal = "Protein Expression and Purification",

publisher = "Elsevier BV",

number = "1",

}

TY - JOUR

T1 - Expression and purification of biologically active IGF-binding proteins using the LCR/Mel expression system

AU - Bagnall, Wendy

AU - Sharpe, Paul M.

AU - Newham, Peter

AU - Tart, Johnathan

AU - Mott, Richard A.

AU - Torr, Vanessa R.

AU - Forder, Robert A.

AU - Needham, M. R C

PY - 2003/1/1

Y1 - 2003/1/1

N2 - The anabolic effects and bioavailability of insulin-like growth factors I and II (IGF-I, IGF-II) are regulated in part by a family of IGF-binding proteins (IGFBPs). There are six known members of the IGFBP family, which share distinct structural characteristics and functional activities. To study the binding properties of these proteins, we have expressed recombinant IGFBP-3 and IGFBP-4 using the LCR/Mel expression system. Using this system, we found that recombinant IGFBP-3 was secreted by Mel cells and had a glycosylation pattern similar to that of native IGFBP-3. Recombinant IGFBP-4 secreted from Mel cells had a molecular size identical to that of non-glycosylated native IGFBP-4. The binding kinetics of recombinant IGFBPs was measured using a solid-phase ligand-binding assay, an in vitro solution-binding assay, and a cellular proliferation assay. IGF-I bound with high affinity to recombinant IGFBP-3 and IGFBP-4 with KDs of

AB - The anabolic effects and bioavailability of insulin-like growth factors I and II (IGF-I, IGF-II) are regulated in part by a family of IGF-binding proteins (IGFBPs). There are six known members of the IGFBP family, which share distinct structural characteristics and functional activities. To study the binding properties of these proteins, we have expressed recombinant IGFBP-3 and IGFBP-4 using the LCR/Mel expression system. Using this system, we found that recombinant IGFBP-3 was secreted by Mel cells and had a glycosylation pattern similar to that of native IGFBP-3. Recombinant IGFBP-4 secreted from Mel cells had a molecular size identical to that of non-glycosylated native IGFBP-4. The binding kinetics of recombinant IGFBPs was measured using a solid-phase ligand-binding assay, an in vitro solution-binding assay, and a cellular proliferation assay. IGF-I bound with high affinity to recombinant IGFBP-3 and IGFBP-4 with KDs of

U2 - 10.1016/S1046-5928(02)00549-1

DO - 10.1016/S1046-5928(02)00549-1

M3 - Article

VL - 27

SP - 1

EP - 11

JO - Protein Expression and Purification

JF - Protein Expression and Purification

IS - 1

ER -

Expression and purification of biologically active IGF-binding proteins using the LCR/Mel expression system

Abstract

Access to Document

Fingerprint

Cite this