A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA

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

doi:https://doi.org/10.1620/tjem.202.155

A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA

Hanawa, Haruo; Watanabe, Ritsuo; Hayashi, Manabu et al.

Tohoku Journal of Experimental Medicine, 2004, 202(3), 155-161

アクセス数:1,474件（2025-07-15 06:08 集計）

固定URL: https://hdl.handle.net/11094/23114

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タイトル	A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA
著者	NRID 1000040282983 1000040282983 著者名 Hanawa, Haruo
	著者名 Watanabe, Ritsuo
	NRID 1000080745787 1000080745787 著者名 Hayashi, Manabu
	NRID 1000040284509 1000040284509 著者名 Yoshida, Tsuyoshi
	著者名 Abe, Satoru
	著者名 Komura, Satoru
	著者名 Liu, Hui
	著者名 Elnaggar, Raafat
	著者名 Chang, He
	NRID 1000090361914 1000090361914 著者名 Okura, Yuji
	NRID 1000000303165 1000000303165 著者名 Kato, Kiminori
	NRID 1000070344447 1000070344447 著者名 Kodama, Makoto
	NRID 1000010293218 1000010293218 著者名 Maruyama, Hiroki
	NRID 1000080229830 1000080229830 著者名 Miyazaki, Junichi
	NRID 1000050143780 1000050143780 著者名 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.
出版者	Tohoku University Medical Press
	東北ジャーナル刊行会
	トウホク　ジャーナル　カンコウカイ
収録物名	Tohoku Journal of Experimental Medicine
巻(号)	202(3)
ページ	155-161
刊行年月	2004-01-09
言語	英語
ハンドルURL	https://hdl.handle.net/11094/23114
PISSN	00408727
NCID	AA00863920
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1620/tjem.202.155
アクセス権	open access
権利情報	© 2004 Tohoku University Medical Press
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	Tohoku Journal of Experimental Medicine. 2004, 202(3), p. 155-161
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
	東北ジャーナル刊行会
	トウホク　ジャーナル　カンコウカイ
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2004-01-09
DC.identifier	https://doi.org/10.1620/tjem.202.155
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
DC.format	23114
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_doi	https://doi.org/10.1620/tjem.202.155
citation_public_url	https://hdl.handle.net/11094/23114

A Novel Method to Assay Proteins in Blood Plasma after Intravenous Injection of Plasmid DNA

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