Journal of Atherosclerosis and Thrombosis
Online ISSN : 1880-3873
Print ISSN : 1340-3478
ISSN-L : 1340-3478
Original Articles
The Generation of Monoclonal Antibodies against Human Peroxisome Proliferator-activated Receptors (PPARs)
Toshiya TanakaTetsu TakenoYuichiro WatanabeYasutoshi UchiyamaTakeshi MurakamiHisahiko YamashitaAkifumi SuzukiRie AoiHiroko IwanariShu-Ying JiangMakoto NaitoKeisuke TachibanaTakefumi DoiAndrew I. ShulmanDavid J. MangelsdorfRaphael ReiterJohan AuwerxTakao HamakuboTatsuhiko Kodama
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2002 Volume 9 Issue 5 Pages 233-242

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Abstract

Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARδ and PPARγ2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence. Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.

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この記事はクリエイティブ・コモンズ [表示 - 非営利 - 継承 4.0 国際]ライセンスの下に提供されています。
https://creativecommons.org/licenses/by-nc-sa/4.0/deed.ja
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