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論文情報
タイトル
A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA
著者
Hanawa, Haruo
Hanawa, Haruo
Watanabe, Ritsuo
Watanabe, Ritsuo
Hayashi, Manabu
Hayashi, Manabu
Yoshida, Tsuyoshi
Yoshida, Tsuyoshi
Abe, Satoru
Abe, Satoru
Komura, Satoru
Komura, Satoru
Liu, Hui
Liu, Hui
Elnaggar, Raafat
Elnaggar, Raafat
Chang, He
Chang, He
Okura, Yuji
Okura, Yuji
Kato, Kiminori
Kato, Kiminori
Kodama, Makoto
Kodama, Makoto
Maruyama, Hiroki
Maruyama, Hiroki
Miyazaki, Junichi
Miyazaki, Junichi
Aizawa, Yoshifusa
Aizawa, Yoshifusa
抄録
Gene therapy is expected to lead to new and useful methods to treat diseases. The development of assays to quantitate gene-therapy-derived proteins circulating in blood will be essential to investigate the effects and side effects of the introduced proteins. The purpose of this study is to evaluate whether a protein circulating at trace concentrations in blood can be measured by tagging a peptide corresponding to glucagon residues 19-29 onto its C-terminal end. We constructed plasmids encoding chimeric proteins and transferred them into rats by hydrodynamics-based delivery. When plasmids encoding human IL8-glucagon 19-29 chimeric protein were injected into rats to evaluate the accuracy of this method, there was a high correlation between chimeric proteins measured by an enzyme-linked immunosorbent assay for human IL8 and one by a radioimmunoassay for glucagon. Furthermore, when plasmids coding rat IFN gamma receptor IgG-Fc glucagon 19-29 chimeric protein were injected to evaluate the time course of chimeric proteins in blood plasma, we could calculate the concentrations in blood from 10 μl plasma samples using glucagon 19-29 tag as follows: 2815±2318 ng/ml after 4 hours (mean±S.D.), 6061±2789 ng/ml after 8 hours, 5752±2270 ng/ml after 12 hours, 2870±1062 ng/ml after one day, 1440±334 ng/ml after three days, 1120±433 ng/ml after seven days, and 281±162 ng/ml after 16 days. Blood sugar levels which might have been increased by glucagon did not increase even at peak chimeric protein concentrations. These results demonstrate a useful and convenient method to assay gene therapy products circulating in blood using a glucagon 19-29 tagging vector.
公開者
Tohoku University Medical Press
公開者の別表記
東北ジャーナル刊行会
公開者 (ヨミ)
トウホク ジャーナル カンコウカイ
掲載誌名
Tohoku Journal of Experimental Medicine
巻
202
号
3
開始ページ
155
終了ページ
161
刊行年月
2004-01-09
ISSN
00408727
NCID
AA00863920
DOI (出版社版)
info:doi/10.1620/tjem.202.155
URL
http://hdl.handle.net/11094/23114
権利情報
© 2004 Tohoku University Medical Press
言語
英語
カテゴリ
学術雑誌論文 Journal Article
論文詳細を表示
著者版フラグ
publisher
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学術雑誌論文
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dcmi資源タイプ
text
DCTERMS.bibliographicCitation
Tohoku Journal of Experimental Medicine.202(3) P.155-P.161
DC.title
A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA
DC.creator
Hanawa, Haruo
Watanabe, Ritsuo
Hayashi, Manabu
Yoshida, Tsuyoshi
Abe, Satoru
Komura, Satoru
Liu, Hui
Elnaggar, Raafat
Chang, He
Okura, Yuji
Kato, Kiminori
Kodama, Makoto
Maruyama, Hiroki
Miyazaki, Junichi
Aizawa, Yoshifusa
DC.publisher
Tohoku University Medical Press
DC.language" scheme="DCTERMS.RFC1766
英語
DCTERMS.issued" scheme="DCTERMS.W3CDTF
2004-01-09
DC.identifier
info:doi/10.1620/tjem.202.155
DC.identifier" scheme="DCTERMS.URI
http://hdl.handle.net/11094/23114
DCTERMS.abstract
Gene therapy is expected to lead to new and useful methods to treat diseases. The development of assays to quantitate gene-therapy-derived proteins circulating in blood will be essential to investigate the effects and side effects of the introduced proteins. The purpose of this study is to evaluate whether a protein circulating at trace concentrations in blood can be measured by tagging a peptide corresponding to glucagon residues 19-29 onto its C-terminal end. We constructed plasmids encoding chimeric proteins and transferred them into rats by hydrodynamics-based delivery. When plasmids encoding human IL8-glucagon 19-29 chimeric protein were injected into rats to evaluate the accuracy of this method, there was a high correlation between chimeric proteins measured by an enzyme-linked immunosorbent assay for human IL8 and one by a radioimmunoassay for glucagon. Furthermore, when plasmids coding rat IFN gamma receptor IgG-Fc glucagon 19-29 chimeric protein were injected to evaluate the time course of chimeric proteins in blood plasma, we could calculate the concentrations in blood from 10 μl plasma samples using glucagon 19-29 tag as follows: 2815±2318 ng/ml after 4 hours (mean±S.D.), 6061±2789 ng/ml after 8 hours, 5752±2270 ng/ml after 12 hours, 2870±1062 ng/ml after one day, 1440±334 ng/ml after three days, 1120±433 ng/ml after seven days, and 281±162 ng/ml after 16 days. Blood sugar levels which might have been increased by glucagon did not increase even at peak chimeric protein concentrations. These results demonstrate a useful and convenient method to assay gene therapy products circulating in blood using a glucagon 19-29 tagging vector.
DC.rights
© 2004 Tohoku University Medical Press
citation_title
A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA
citation_author
Hanawa, Haruo
Watanabe, Ritsuo
Hayashi, Manabu
Yoshida, Tsuyoshi
Abe, Satoru
Komura, Satoru
Liu, Hui
Elnaggar, Raafat
Chang, He
Okura, Yuji
Kato, Kiminori
Kodama, Makoto
Maruyama, Hiroki
Miyazaki, Junichi
Aizawa, Yoshifusa
citation_publisher
Tohoku University Medical Press
citation_language
英語
citation_date
2004-01-09
citation_journal_title
Tohoku Journal of Experimental Medicine
citation_volume
202
citation_issue
3
citation_firstpage
155
citation_lastpage
161
citation_issn
00408727
citation_doi
info:doi/10.1620/tjem.202.155
citation_public_url
http://hdl.handle.net/11094/23114