title: Generation of monoclonal antibody recognized by the GXXXG motif (glycine zipper) of prion protein.
abstract: To develop monoclonal antibodies (MAbs) to react with normal prion protein (PrPC) and abnormal isoform of prion protein (PrPSc), PrPSc was isolated from brains of 263 K scrapie-infected hamsters and immunized to PrP knockout mice. We developed two hybridomas, 3F10 and 1C5 (IgG1), of which epitope mappings were screened by using glutathione S-transferase (GST) fusion proteins of recombinant hamster prion protein and suitable peptides. 3F10 showed a high affinity for hamster and mouse PrP and was demonstrated to recognize the residues 137-151. 1C5 recognizes the region 119-130 corresponding to the GXXXG motif, the glycine zipper region, conserved in all mammals. In the immunohistochemical analysis, the positive staining for PrPSc was observed in the extracellular compartment of scrapie-infected brains but not in the normal brains. However, in Western blot, these antibodies recognized both normal and abnormal prion proteins. These results suggested that the developed mouse MAbs are specific to prion protein and can recognize abnormal prion protein more effectively than normal prion protein in immunohistochemistry. Therefore, these antibodies could be utilized as a useful reagent for the analysis of biochemical, structural, and functional properties between PrPC and PrPSc.
authors.affiliation: Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
authors.forename: Jin-Kyu
authors.initials: JK
authors.lastname: Choi
authors.forename: Seok-Ju
authors.initials: SJ
authors.lastname: Park
authors.forename: Yong-Chul
authors.initials: YC
authors.lastname: Jun
authors.forename: Jae-Min
authors.initials: JM
authors.lastname: Oh
authors.forename: Byung-Hoon
authors.initials: BH
authors.lastname: Jeong
authors.forename: Hyun-Pil
authors.initials: HP
authors.lastname: Lee
authors.forename: Sue-Nie
authors.initials: SN
authors.lastname: Park
authors.forename: Richard I
authors.initials: RI
authors.lastname: Carp
authors.forename: Yong-Sun
authors.initials: YS
authors.lastname: Kim
mesh_terms: D020816:Amino Acid Motifs / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism*; D015153:Blotting, Western; D006224:Cricetinae; D018604:Epitope Mapping; D005260:Female; D006825:Hybridomas / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D018345:Mice, Knockout; D018621:PrPC Proteins / Q000235:genetics; D011328:Prions / Q000737:chemistry* / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D011328:Prions