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

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Title: Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity
Authors: Nakano, Keiichi Browse this author
Tamura, Shogo Browse this author
Otuka, Kohei Browse this author
Niizeki, Noriyasu Browse this author
Shigemura, Masahiko Browse this author
Shimizu, Chikara Browse this author
Matsuno, Kazuhiko Browse this author
Kobayashi, Seiichi Browse this author
Moriyama, Takanori Browse this author →KAKEN DB
Keywords: Three-dimensional gel electrophoresis
Monoclonal proteins
Issue Date: 15-Jul-2013
Journal Title: Analytical Biochemistry
Volume: 438
Issue: 2
Start Page: 117
End Page: 123
Publisher DOI: 10.1016/j.ab.2013.03.013
PMID: 23541520
Abstract: 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.
Type: article (author version)
Appears in Collections:北海道大学病院 (Hokkaido University Hospital) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 中野 恵一

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