title: Viral envelope protein gp64 transgenic mouse facilitates the generation of monoclonal antibodies against exogenous membrane proteins displayed on baculovirus.
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.
authors: Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co. Ltd., 1-135 Komakado, Gotemba, Shizuoka, 412-8513, Japan.
authors: Ryoichi
authors: R
authors: Saitoh
authors: Toshihiko
authors: T
authors: Ohtomo
authors: Yoshiki
authors: Y
authors: Yamada
authors: Nobuo
authors: N
authors: Kamada
authors: Jun-Ichi
authors: J
authors: Nezu
authors: Naoki
authors: N
authors: Kimura
authors: Shin-Ichi
authors: S
authors: Funahashi
authors: Koh
authors: K
authors: Furugaki
authors: Takeshi
authors: T
authors: Yoshino
authors: Yosuke
authors: Y
authors: Kawase
authors: Atsuhiko
authors: A
authors: Kato
authors: Otoya
authors: O
authors: Ueda
authors: Kou-Ichi
authors: K
authors: Jishage
authors: Masami
authors: M
authors: Suzuki
authors: Rie
authors: R
authors: Fukuda
authors: Masahiro
authors: M
authors: Arai
authors: Hiroko
authors: H
authors: Iwanari
authors: Kazuaki
authors: K
authors: Takahashi
authors: Toshiko
authors: T
authors: Sakihama
authors: Iwao
authors: I
authors: Ohizumi
authors: Tatsuhiko
authors: T
authors: Kodama
authors: Masayuki
authors: M
authors: Tsuchiya
authors: Takao
authors: T
authors: Hamakubo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D016466:CHO Cells; D015815:Cell Adhesion Molecules / Q000235:genetics*; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D007114:Immunization; D008562:Membrane Glycoproteins / Q000235:genetics*; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008822:Mice, Transgenic; D019151:Peptide Library; D000074011:Peptide Transporter 1; D054390:Receptors, CCR2; D019707:Receptors, Chemokine / Q000276:immunology; D027981:Symporters / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; C515335:Ccr2 protein, mouse; D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019151:Peptide Library; D000074011:Peptide Transporter 1; D054390:Receptors, CCR2; D019707:Receptors, Chemokine; D027981:Symporters; D014759:Viral Envelope Proteins; D014764:Viral Proteins