Preparation of monoclonal antibodies against prion proteins with full-length hamster PrP.
OBJECTIVE
To prepare the PrP specific monoclonal antibodies (mAbs) that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases.


METHODS
Several BALB/c mice were immunized with recombinant hamster prion protein (HaPrP). Three hybridoma cell lines designated as B7, B9, and B10, secreting monoclonal antibodies against HaPrP, were established by hybridoma technique. The mAbs reactivities were evaluated with ELISA, Western blot, and immunohistochemistry.


RESULTS
The mAbs produced by these cell lines reacted well with different recombinant hamster PrP proteins. Western blot analyses showed that mAbs B7 and B9 reacted with PrPSc from the scrapie-infected animals after proteinase K digestion with three glycosylated forms. The mAbs exhibited cross-reactivity with various PrPC from several other mammalian species, including humans and cattles. Immunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPsSc.


CONCLUSIONS
The mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases.
Xiao
Xin-Li
XL
School of Medicine, Xi'an Jiao-Tong University, Shaanxi, China.
Jiang
Hui-Ying
HY
Zhang
Jin
J
Han
Jun
J
Nie
Kai
K
Zhou
Xiao-Bo
XB
Huang
Yin-Xia
YX
Chen
Lan
L
Zhou
Wei
W
Zhang
Bao-Yun
BY
Liu
Yong
Y
Dong
Xiao-Ping
XP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007114:Immunization; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D018621:PrPC Proteins / Q000235:genetics / Q000276:immunology*; D018620:PrPSc Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011994:Recombinant Proteins