Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein

Enzyme-linked immunosorbent assay (ELISA) performed using extensively purified bacterially expressed bovine prion protein (PrP) shows decreased cross-reactivity. We generated a transduced Madin–Darby bovine kidney (MDBK) cell line continuously expressing glycosylphosphatidylinositol (GPI)-anchorless...

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Autores principales: KIM, Hyo-Jin, ROH, In-Soon, PARK, Hoo-Chang, AHN, Su Bi, SUH, Tae-Young, PARK, Kyung-Je, KANG, Hae-Eun, SOHN, Hyun-Joo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2018
Materias:
Virology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6021889/
https://www.ncbi.nlm.nih.gov/pubmed/29618668
http://dx.doi.org/10.1292/jvms.17-0521
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author KIM, Hyo-Jin
ROH, In-Soon
PARK, Hoo-Chang
AHN, Su Bi
SUH, Tae-Young
PARK, Kyung-Je
KANG, Hae-Eun
SOHN, Hyun-Joo
author_facet KIM, Hyo-Jin
ROH, In-Soon
PARK, Hoo-Chang
AHN, Su Bi
SUH, Tae-Young
PARK, Kyung-Je
KANG, Hae-Eun
SOHN, Hyun-Joo
author_sort KIM, Hyo-Jin
collection PubMed
description Enzyme-linked immunosorbent assay (ELISA) performed using extensively purified bacterially expressed bovine prion protein (PrP) shows decreased cross-reactivity. We generated a transduced Madin–Darby bovine kidney (MDBK) cell line continuously expressing glycosylphosphatidylinositol (GPI)-anchorless bovine PrP (designated as MDBK ∆GPI protein) by using a lentiviral expression system. The present study also described the method for purifying bovine PrP through sequential culturing without the need for complex purification protocol. Our results showed that the purified bovine PrP could be used as an immunogen for developing anti-PrP monoclonal antibodies. Together, our results suggest that the new GPI-anchorless bovine PrP and its purification method can be used for performing basic studies for employing a cell-based approach.
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spelling pubmed-60218892018-07-06 Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein KIM, Hyo-Jin ROH, In-Soon PARK, Hoo-Chang AHN, Su Bi SUH, Tae-Young PARK, Kyung-Je KANG, Hae-Eun SOHN, Hyun-Joo J Vet Med Sci Virology Enzyme-linked immunosorbent assay (ELISA) performed using extensively purified bacterially expressed bovine prion protein (PrP) shows decreased cross-reactivity. We generated a transduced Madin–Darby bovine kidney (MDBK) cell line continuously expressing glycosylphosphatidylinositol (GPI)-anchorless bovine PrP (designated as MDBK ∆GPI protein) by using a lentiviral expression system. The present study also described the method for purifying bovine PrP through sequential culturing without the need for complex purification protocol. Our results showed that the purified bovine PrP could be used as an immunogen for developing anti-PrP monoclonal antibodies. Together, our results suggest that the new GPI-anchorless bovine PrP and its purification method can be used for performing basic studies for employing a cell-based approach. The Japanese Society of Veterinary Science 2018-04-04 2018-06 /pmc/articles/PMC6021889/ /pubmed/29618668 http://dx.doi.org/10.1292/jvms.17-0521 Text en ©2018 The Japanese Society of Veterinary Science This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/)
spellingShingle Virology
KIM, Hyo-Jin
ROH, In-Soon
PARK, Hoo-Chang
AHN, Su Bi
SUH, Tae-Young
PARK, Kyung-Je
KANG, Hae-Eun
SOHN, Hyun-Joo
Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein
title Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein
title_full Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein
title_fullStr Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein
title_full_unstemmed Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein
title_short Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein
title_sort establishment of a madin–darby bovine kidney cell line expressing anchorless bovine prion protein
topic Virology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6021889/
https://www.ncbi.nlm.nih.gov/pubmed/29618668
http://dx.doi.org/10.1292/jvms.17-0521
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