title: Characterization of stress-exposed granulocyte colony stimulating factor using ELISA and hydrogen/deuterium exchange mass spectrometry.
abstract: Information on the higher-order structure is important in the development of biopharmaceutical drugs. Recently, hydrogen/deuterium exchange coupled with mass spectrometry (HDX-MS) has been widely used as a tool to evaluate protein conformation, and unique automated systems for HDX-MS are now commercially available. To investigate the potential of this technique for the prediction of the activity of biopharmaceuticals, granulocyte colony stimulating factor (G-CSF), which had been subjected to three different stress types, was analyzed using HDX-MS and through comparison with receptor-binding activity. It was found that HDX-MS, in combination with ion mobility separation, was able to identify conformational changes in G-CSF induced by stress, and a good correlation with the receptor-binding activity was demonstrated, which cannot be completely determined by conventional peptide mapping alone. The direct evaluation of biological activity using bioassay is absolutely imperative in biopharmaceutical development, but HDX-MS can provide the alternative information in a short time on the extent and location of the structural damage caused by stresses. Furthermore, the present study suggests the possibility of this system being a versatile evaluation method for the preservation stability of biopharmaceuticals.
authors: Bio Process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., Takasaki, Gunma, 370-0013, Japan, daisuke.tsuchida@kyowa-kirin.co.jp.
authors: Daisuke
authors: D
authors: Tsuchida
authors: Katsuyoshi
authors: K
authors: Yamazaki
authors: Satoko
authors: S
authors: Akashi
mesh_terms: D000595:Amino Acid Sequence; D041961:Deuterium Exchange Measurement / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor