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Title: Proteoliposome-based Selection of a Recombinant Antibody Fragment Against the Human M2 Muscarinic Acetylcholine Receptor.
Authors: Suharni
Nomura, Yayoi
Arakawa, Takatoshi
Hino, Tomoya
Abe, Hitomi
Nakada-Nakura, Yoshiko
Sato, Yumi
Iwanari, Hiroko
Shiroishi, Mitsunori
Asada, Hidetsugu  kyouindb  KAKEN_id  orcid https://orcid.org/0000-0001-6255-4728 (unconfirmed)
Shimamura, Tatsuro  kyouindb  KAKEN_id
Murata, Takeshi
Kobayashi, Takuya
Hamakubo, Takao
Iwata, So  kyouindb  KAKEN_id
Nomura, Norimichi  kyouindb  KAKEN_id
Author's alias: 野村, 紀通
Issue Date: 29-Dec-2014
Publisher: Mary Ann Liebert, Inc.
Journal title: Monoclonal antibodies in immunodiagnosis and immunotherapy
Volume: 33
Issue: 6
Start page: 378
End page: 385
Abstract: The development of antibodies against human G-protein-coupled receptors (GPCRs) has achieved limited success, which has mainly been attributed to their low stability in a detergent-solubilized state. We herein describe a method that can generally be applied to the selection of phage display libraries with human GPCRs reconstituted in liposomes. A key feature of this approach is the production of biotinylated proteoliposomes that can be immobilized on the surface of streptavidin-coupled microplates or paramagnetic beads and used as a binding target for antibodies. As an example, we isolated a single chain Fv fragment from an immune phage library that specifically binds to the human M2 muscarinic acetylcholine receptor with nanomolar affinity. The selected antibody fragment recognized the GPCR in both detergent-solubilized and membrane-embedded forms, which suggests that it may be a potentially valuable tool for structural and functional studies of the GPCR. The use of proteoliposomes as immunogens and screening bait will facilitate the application of phage display to this difficult class of membrane proteins.
Rights: The final published version is available from Mary Ann Liebert, Inc., publishers at http://dx.doi.org/10.1089/mab.2014.0041.
この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。This is not the published version. Please cite only the published version.
URI: http://hdl.handle.net/2433/193246
DOI(Published Version): 10.1089/mab.2014.0041
PubMed ID: 25545206
Appears in Collections:Journal Articles

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