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Aqueous Stability of Human Epidermal Growth Factor 1-48

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Abstract

Human epidermal growth factor 1-48 (hEGF 1-48, Des(49-53)hEGF) is a single chain polypeptide (48 amino acids; 3 disulfide bonds; 5445 Da) possessing a broad spectrum of biologic activity including the stimulation of cell proliferation and tissue growth. In this study, three primary aqueous degradation products of hEGF 1-48 were isolated using isocratic, reverse phase/ion-pair HPLC. The degradation products were characterized using amino acid sequencing, electrospray ionization mass spectrometry, isoelectric focusing, and degradation kinetics. Results indicate that hEGF 1-48 degrades via oxidation (Met21), deamidation (Asn1), and succinimide formation (Asp11). The relative contribution of each degradation pathway to the overall stability of hEGF 1-48 changes as a function of solution pH and storage condition. Succinimide formation at Asp11 is favored at pH < 6 in which aspartic acid is present mostly in its protonated form. Deamidation of Asn1 is favored at pH > 6. The relative contribution of Met21 oxidation is increased with decreasing temperature, storage as a frozen solution (−20°C), and exposure to fluorescent light.

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Author notes

  1. Richard I. Senderoff

    Present address: ZymoGenetics, Inc., 4225 Roosevelt Way N.E., Seattle, Washington, 98105

Authors and Affiliations

  1. Warner-Lambert Company, Morris Plains, New Jersey, 07950

    Richard I. Senderoff, Scott C. Wootton, Amal M. Boctor, Tsun Ming Chen, Thomas N. Julian & Galen W. Radebaugh

  2. Department of Chemistry, Parke-Davis Pharmaceutical Research Division, Warner-Lambert Company, Ann Arbor, Michigan, 48105

    Anne B. Giordani

Authors
  1. Richard I. Senderoff
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  2. Scott C. Wootton
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  3. Amal M. Boctor
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  5. Anne B. Giordani
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Senderoff, R.I., Wootton, S.C., Boctor, A.M. et al. Aqueous Stability of Human Epidermal Growth Factor 1-48. Pharm Res 11, 1712–1720 (1994). https://doi.org/10.1023/A:1018903014204

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