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Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells
BACKGROUND: A hallmark of prion disease is the transformation of normal cellular prion protein (PrPc) into an infectious disease-associated isoform, (PrPsc). Anti-prion protein monoclonal antibodies are invaluable for structure-function studies of PrP molecules. Furthermore recent in vitro and in vi...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2007
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1933425/ https://www.ncbi.nlm.nih.gov/pubmed/17605808 http://dx.doi.org/10.1186/1472-6750-7-38 |
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author | Flego, Michela Ascione, Alessandro Zamboni, Silvia Dupuis, Maria L Imperiale, Valentina Cianfriglia, Maurizio |
author_facet | Flego, Michela Ascione, Alessandro Zamboni, Silvia Dupuis, Maria L Imperiale, Valentina Cianfriglia, Maurizio |
author_sort | Flego, Michela |
collection | PubMed |
description | BACKGROUND: A hallmark of prion disease is the transformation of normal cellular prion protein (PrPc) into an infectious disease-associated isoform, (PrPsc). Anti-prion protein monoclonal antibodies are invaluable for structure-function studies of PrP molecules. Furthermore recent in vitro and in vivo studies indicate that anti-PrP monoclonal antibodies can prevent the incorporation of PrPc into propagating prions. In the present article, we show two new human phage antibodies, isolated on recombinant hamster prion protein (rHaPrP). RESULTS: We adopted an antibody phage display strategy to isolate specific human antibodies directed towards rHaPrP which has been used as a bait for panning the synthetic ETH-2 antibody phage library. Two phage antibodies clones named MA3.B4 and MA3.G3 were isolated and characterized under genetic biochemical and immunocytochemical aspects. The clones were found to recognize the prion protein in ELISA studies. In flow-cytometry studies, these human single chain Fragment variable (scFv) phage-antibodies show a well defined pattern of reactivity on human lymphoblastoid and myeloid cells. CONCLUSION: Sequence analysis of the gene encoding for the antibody fragments and antigen recognition patterns determined by flow-cytometry analysis indicate that the isolated scFvs recognize novel epitopes in the PrPc molecule. These new anti PrPc human antibodies are unique reagents for prion protein detection and may represent a biologic platform to develop new reagents to treat PrPsc associated disease. |
format | Text |
id | pubmed-1933425 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-19334252007-07-26 Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells Flego, Michela Ascione, Alessandro Zamboni, Silvia Dupuis, Maria L Imperiale, Valentina Cianfriglia, Maurizio BMC Biotechnol Research Article BACKGROUND: A hallmark of prion disease is the transformation of normal cellular prion protein (PrPc) into an infectious disease-associated isoform, (PrPsc). Anti-prion protein monoclonal antibodies are invaluable for structure-function studies of PrP molecules. Furthermore recent in vitro and in vivo studies indicate that anti-PrP monoclonal antibodies can prevent the incorporation of PrPc into propagating prions. In the present article, we show two new human phage antibodies, isolated on recombinant hamster prion protein (rHaPrP). RESULTS: We adopted an antibody phage display strategy to isolate specific human antibodies directed towards rHaPrP which has been used as a bait for panning the synthetic ETH-2 antibody phage library. Two phage antibodies clones named MA3.B4 and MA3.G3 were isolated and characterized under genetic biochemical and immunocytochemical aspects. The clones were found to recognize the prion protein in ELISA studies. In flow-cytometry studies, these human single chain Fragment variable (scFv) phage-antibodies show a well defined pattern of reactivity on human lymphoblastoid and myeloid cells. CONCLUSION: Sequence analysis of the gene encoding for the antibody fragments and antigen recognition patterns determined by flow-cytometry analysis indicate that the isolated scFvs recognize novel epitopes in the PrPc molecule. These new anti PrPc human antibodies are unique reagents for prion protein detection and may represent a biologic platform to develop new reagents to treat PrPsc associated disease. BioMed Central 2007-07-02 /pmc/articles/PMC1933425/ /pubmed/17605808 http://dx.doi.org/10.1186/1472-6750-7-38 Text en Copyright © 2007 Flego et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Flego, Michela Ascione, Alessandro Zamboni, Silvia Dupuis, Maria L Imperiale, Valentina Cianfriglia, Maurizio Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
title | Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
title_full | Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
title_fullStr | Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
title_full_unstemmed | Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
title_short | Generation of human scFvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
title_sort | generation of human scfvs antibodies recognizing a prion protein epitope expressed on the surface of human lymphoblastoid cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1933425/ https://www.ncbi.nlm.nih.gov/pubmed/17605808 http://dx.doi.org/10.1186/1472-6750-7-38 |
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