Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity

Nakano, Keiichi; Tamura, Shogo; Otuka, Kohei; Niizeki, Noriyasu; Shigemura, Masahiko; Shimizu, Chikara; Matsuno, Kazuhiko; Kobayashi, Seiichi; Moriyama, Takanori

Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity

Nakano, Keiichi; Tamura, Shogo; Otuka, Kohei et al.

Analytical Biochemistry, 2013, 438(2), 117-123

Permalink : https://hdl.handle.net/2115/53040

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アクセス権	open access
URI	https://hdl.handle.net/2115/53040
タイトル	Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity
著者	Nakano, Keiichi ; Tamura, Shogo ; Otuka, Kohei ; Niizeki, Noriyasu ; Shigemura, Masahiko ; Shimizu, Chikara ; Matsuno, Kazuhiko ; Kobayashi, Seiichi ; Moriyama, Takanori
著者名 Nakano, Keiichi
著者名 Tamura, Shogo
著者名 Otuka, Kohei
著者名 Niizeki, Noriyasu
著者名 Shigemura, Masahiko
著者名 Shimizu, Chikara
著者名 Matsuno, Kazuhiko
著者名 Kobayashi, Seiichi
e-Rad_Researcher 20312423 著者名 Moriyama, Takanori
言語	英語
キーワード	Three-dimensional gel electrophoresis
	Monoclonal proteins
	Heterogeneity
発行日	2013-07-15
収録物名	Analytical Biochemistry
巻(号)	438(2)
ページ	117-123
抄録	Three-dimensional gel electrophoresis (3-DE), which combines agarose gel electrophoresis and isoelectric focusing/SDS–PAGE, was developed to characterize monoclonal proteins (M-proteins). However, the original 3-DE method has not been optimized and its specificity has not been demonstrated. The main goal of this study was to optimize the 3-DE procedure and then compare it with 2-DE. We developed a highly sensitive 3-DE method in which M-proteins are extracted from a first-dimension agarose gel, by diffusing into 150 mM NaCl, and the recovery of M-proteins was 90.6%. To validate the utility of the highly sensitive 3-DE, we compared it with the original 3-DE method. We found that highly sensitive 3-DE provided for greater M-protein recovery and was more effective in terms of detecting spots on SDS–PAGE gels than the original 3-DE. Moreover, highly sensitive 3-DE separates residual normal IgG from M-proteins, which could not be done by 2-DE. Applying the highly sensitive 3-DE to clinical samples, we found that the characteristics of M-proteins vary tremendously between individuals. We believe that our highly sensitive 3-DE method described here will prove useful in further studies of the heterogeneity of M-proteins.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
PISSN	0003-2697
関連情報	PMID 23541520
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.ab.2013.03.013