TY - JOUR
T1 - A novel anti-prion protein monoclonal antibody and its single-chain fragment variable derivative with ability to inhibit abnormal prion protein accumulation in cultured cells
AU - Shimizu, Yoshihisa
AU - Kaku-Ushiki, Yuko
AU - Iwamaru, Yoshifumi
AU - Muramoto, Tamaki
AU - Kitamoto, Tetsuyuki
AU - Yokoyama, Takashi
AU - Mohri, Shirou
AU - Tagawa, Yuichi
N1 - Copyright:
Copyright 2010 Elsevier B.V., All rights reserved.
PY - 2010/2
Y1 - 2010/2
N2 - mAbs T1 and T2 were established by immunizing PrP gene ablated mice with recombinant MoPrP of residues 121-231. Both mAbs were cross-reactive with PrP fromhamster, sheep, cattle and deer. A linear epitope of mAb T1 was identified at residues 137-143 of MoPrP and buried in PrPC expressed on the cell surface. mAb T1 showed no inhibitory effect on accumulation of PrPSc in cultured scrapie-infected neuroblastoma (ScN2a) cells. In contrast, mAb T2 recognized a discontinuous epitope ranged on, or structured by, residues 132-217 and this epitope was exposed on the cell surface PrPC.mAb T2 showed an excellent inhibitory effect onPrPSc accumulation in vitro at a50%inhibitory concentration of 0.02 μg/ml (0.14 nM). The scFv form of mAb T2 (scFv T2) was secreted in neuroblastoma (N2a58) cell cultures by transfection through eukaryotic secretion vector. Coculturing of ScN2a cells with scFv T2-producing N2a58 cells induced a clear inhibitory effect on PrPSc accumulation, suggesting that scFv T2 could potentially be an immunotherapeutic tool for prion diseases by inhibition of PrPSc accumulation.
AB - mAbs T1 and T2 were established by immunizing PrP gene ablated mice with recombinant MoPrP of residues 121-231. Both mAbs were cross-reactive with PrP fromhamster, sheep, cattle and deer. A linear epitope of mAb T1 was identified at residues 137-143 of MoPrP and buried in PrPC expressed on the cell surface. mAb T1 showed no inhibitory effect on accumulation of PrPSc in cultured scrapie-infected neuroblastoma (ScN2a) cells. In contrast, mAb T2 recognized a discontinuous epitope ranged on, or structured by, residues 132-217 and this epitope was exposed on the cell surface PrPC.mAb T2 showed an excellent inhibitory effect onPrPSc accumulation in vitro at a50%inhibitory concentration of 0.02 μg/ml (0.14 nM). The scFv form of mAb T2 (scFv T2) was secreted in neuroblastoma (N2a58) cell cultures by transfection through eukaryotic secretion vector. Coculturing of ScN2a cells with scFv T2-producing N2a58 cells induced a clear inhibitory effect on PrPSc accumulation, suggesting that scFv T2 could potentially be an immunotherapeutic tool for prion diseases by inhibition of PrPSc accumulation.
KW - Anti-prion effect
KW - Monoclonal antibody
KW - Single-chain fragment variable region
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U2 - 10.1111/j.1348-0421.2009.00190.x
DO - 10.1111/j.1348-0421.2009.00190.x
M3 - Article
C2 - 20377745
AN - SCOPUS:76649106715
VL - 54
SP - 112
EP - 121
JO - Microbiology and Immunology
JF - Microbiology and Immunology
SN - 0385-5600
IS - 2
ER -