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Application of Horseradish Peroxidase-Encapsulated Liposomes as Labels for Immunodotblotting

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Title: Application of Horseradish Peroxidase-Encapsulated Liposomes as Labels for Immunodotblotting
Authors: SUITA, Takahiro Browse this author
TANI, Hirofumi Browse this author →KAKEN DB
KAMIDATE, Tamio Browse this author
Issue Date: May-2000
Publisher: 日本分析化学会
Journal Title: Analytical Sciences
Volume: 16
Issue: 5
Start Page: 527
End Page: 529
Publisher DOI: 10.2116/analsci.16.527
Abstract: Liposomes are spherical vesicles consisting of phospholipid bilayers surrounding an aqueous volume. Recently, attention has focused on liposomes as a signal-enhancement agent, since a number of marker molecules can be encapsulated in their aqueous interior. An antibody labeled with liposome encapsulating enzyme, such as glucose oxidase, has been employed for immunoassays. On the other hand, HRP-conjugated liposomes were prepared by covalently attaching HRP to the outside of the lipid bilayer of liposome. However, the preparation of HRP-conjugated liposomes was tedious and time-consuming, since several reaction steps are required for linking HRP covalently to the surface of liposomes. In order to hold simply HRP in liposomes, we examined how to encapsulate HRP into the aqueous interior of liposomes prepared by an extrusion technique (VETs).4 In addition, the HRP-encapsulated VETs were coupled covalently to anti-rabbit IgG(antibodytagged VETs).5 the detection of HRP encapsulated in the antibody-tagged VETs was made by luminol chemiluminescent (CL) method. On the other hand, biotin-tagged molecules are currently more universal as a marker in immunoassays compared to antibody-tagged marker moleucles. In the present study, the HRP-encapsulated liposomes containing biotinylated dipalmitoylphosphatidylethanolamin were prepared by an extrusion technique (biotin-tagged VETs). In addition, biotin-tagged VETs were applied to labels in immunodotblotting of rabbit IgG.
Type: article
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 谷 博文

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