Further purification of epidermal growth factor by high-performance liquid chromatography

Lynn M. Matrisian; Brent R. Larsen; Joanne S. Finch; Bruce E. Magun

doi:10.1016/0003-2697(82)90015-X

Further purification of epidermal growth factor by high-performance liquid chromatography

Lynn M. Matrisian, Brent R. Larsen, Joanne S. Finch, Bruce E. Magun

Knight Cancer Biology Program

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

56 Scopus citations

Abstract

Epidermal growth factor (EGF) purified by the method of Savage and Cohen (J. Biol. Chem. 247, 7601-7611 (1972) using DEAE-cellulose chromatography as the final purification step was further resolved into two major uv-absorbing components by reverse-phase high-performance liquid chromatography (HPLC). Both components, referred to as α-EGF and β-EGF, competed with ¹²⁵I-labeled EGF for the EGF receptor, induced premature eye opening in neonatal mice, and had an amino acid composition similar to that published by Savage et al. (J. Biol. Chem.247, 7612-7621 (1972). β-EGF migrated slightly faster than α-EGF during sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. α-EGF was fourfold more potent than β-EGF and was 10-fold more potent than DEAE-purified EGF in stimulating DNA synthesis in quiescent Rat-1 cells. HPLC purification of EGF can replace the DEAE-cellulose chromatography step currently used and produces a more potent and less heterogeneous EGF species.

Original language	English (US)
Pages (from-to)	339-351
Number of pages	13
Journal	Analytical Biochemistry
Volume	125
Issue number	2
DOIs	https://doi.org/10.1016/0003-2697(82)90015-X
State	Published - Sep 15 1982

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Further purification of epidermal growth factor by high-performance liquid chromatography",

abstract = "Epidermal growth factor (EGF) purified by the method of Savage and Cohen (J. Biol. Chem. 247, 7601-7611 (1972) using DEAE-cellulose chromatography as the final purification step was further resolved into two major uv-absorbing components by reverse-phase high-performance liquid chromatography (HPLC). Both components, referred to as α-EGF and β-EGF, competed with 125I-labeled EGF for the EGF receptor, induced premature eye opening in neonatal mice, and had an amino acid composition similar to that published by Savage et al. (J. Biol. Chem.247, 7612-7621 (1972). β-EGF migrated slightly faster than α-EGF during sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. α-EGF was fourfold more potent than β-EGF and was 10-fold more potent than DEAE-purified EGF in stimulating DNA synthesis in quiescent Rat-1 cells. HPLC purification of EGF can replace the DEAE-cellulose chromatography step currently used and produces a more potent and less heterogeneous EGF species.",

author = "Matrisian, {Lynn M.} and Larsen, {Brent R.} and Finch, {Joanne S.} and Magun, {Bruce E.}",

note = "Funding Information: The authors acknowledge the excellent technical assistance of Chris Fennie and Herb Wagner and secretarial assistance of Vondal Sandum. We thank Dr. Irwin Flink, Dr. Victor Hruby, and Dr. Tom Sawyer for their assistance with the amino acid analyses. We would also like to thank Dr. Stephen Planck for his advice and many helpful discussions. This work was supported by NIH Grant 1 ROI CA29290-01.",

year = "1982",

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doi = "10.1016/0003-2697(82)90015-X",

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AU - Larsen, Brent R.

AU - Finch, Joanne S.

AU - Magun, Bruce E.

N1 - Funding Information: The authors acknowledge the excellent technical assistance of Chris Fennie and Herb Wagner and secretarial assistance of Vondal Sandum. We thank Dr. Irwin Flink, Dr. Victor Hruby, and Dr. Tom Sawyer for their assistance with the amino acid analyses. We would also like to thank Dr. Stephen Planck for his advice and many helpful discussions. This work was supported by NIH Grant 1 ROI CA29290-01.

PY - 1982/9/15

Y1 - 1982/9/15

N2 - Epidermal growth factor (EGF) purified by the method of Savage and Cohen (J. Biol. Chem. 247, 7601-7611 (1972) using DEAE-cellulose chromatography as the final purification step was further resolved into two major uv-absorbing components by reverse-phase high-performance liquid chromatography (HPLC). Both components, referred to as α-EGF and β-EGF, competed with 125I-labeled EGF for the EGF receptor, induced premature eye opening in neonatal mice, and had an amino acid composition similar to that published by Savage et al. (J. Biol. Chem.247, 7612-7621 (1972). β-EGF migrated slightly faster than α-EGF during sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. α-EGF was fourfold more potent than β-EGF and was 10-fold more potent than DEAE-purified EGF in stimulating DNA synthesis in quiescent Rat-1 cells. HPLC purification of EGF can replace the DEAE-cellulose chromatography step currently used and produces a more potent and less heterogeneous EGF species.

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Further purification of epidermal growth factor by high-performance liquid chromatography

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