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Published Online: 17 October 2006

Generation of Monoclonal Antibody Recognized by the GXXXG Motif (Glycine Zipper) of Prion Protein

Publication: Hybridoma
Volume 25, Issue Number 5

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 263K 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.

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cover image Hybridoma
Hybridoma
Volume 25Issue Number 5October 2006
Pages: 271 - 277
PubMed: 17044782

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Published online: 17 October 2006
Published in print: October 2006

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Jin-Kyu Choi
Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
Seok-Ju Park
Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
Yong-Chul Jun
Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
Jae-Min Oh
Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
Byung-Hoon Jeong
Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
Hyun-Pil Lee
Ilsong Institute of Life Science, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.
Sue-Nie Park
National Institute of Toxicological Research, Korea Food and Drug Administration, Seoul, Republic of Korea.
Richard I. Carp
New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York.
Yong-Sun Kim, M.D., Ph.D.
Ilsong Institute of Life Science and Department of Microbiology, College of Medicine, Hallym University, Kyeonggi-Do, Republic of Korea.

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