| Literature DB >> 17374538 |
Ryoichi Saitoh1, Toshihiko Ohtomo, Yoshiki Yamada, Nobuo Kamada, Jun-Ichi Nezu, Naoki Kimura, Shin-Ichi Funahashi, Koh Furugaki, Takeshi Yoshino, Yosuke Kawase, Atsuhiko Kato, Otoya Ueda, Kou-Ichi Jishage, Masami Suzuki, Rie Fukuda, Masahiro Arai, Hiroko Iwanari, Kazuaki Takahashi, Toshiko Sakihama, Iwao Ohizumi, Tatsuhiko Kodama, Masayuki Tsuchiya, Takao Hamakubo.
Abstract
We have been investigating the functional display of multipass membrane protein such as transporter or G-protein coupled receptor on the budded baculovirus (BV). We tested the use of a viral envelope protein gp64 transgenic mouse for the direct immunization of these membrane proteins displayed on BVs. The gp64 transgenic mice showed only a weak response to virus compared to wild type BALB/c mice. Immunizing gp64 transgenic mice with the BV expressing peptide transporter PepT1, we obtained 47 monoclonal antibodies (mAbs). These mAbs were specific to the PepT1 on the pancreatic cancer cells AsPC-1 by fluorocytometric analysis and exhibited antibody-dependent cellular cytotoxicity or complement-dependent cytotoxicity to AsPC-1. We also generated 7 mAbs by immunizing gp64 transgenic mice on a CCR2-deficient background with the BV expressing chemokine receptor CCR2 together with partially purified CCR2. These mAbs possessed specific binding to CCR2 in CHO cells on fluorocytometric analysis, and exhibited neutralizing activities for ligand-dependent inhibition of cyclic AMP production. This method provides a powerful tool for the generation of therapeutic/diagnostic mAbs against membrane proteins.Entities:
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Year: 2007 PMID： 17374538 DOI： 10.1016/j.jim.2007.02.005
Source DB: PubMed Journal: J Immunol Methods ISSN： 0022-1759 Impact factor: 2.303