Mapping of B cell epitopes in measles virus nucleocapsid protein |
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Authors: | A Zvirbliene I Kucinskaite I Sezaite D Samuel K Sasnauskas |
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Institution: | (1) Institute of Biotechnology, Vilnius, Lithuania;(2) Enteric, Respiratory and Neurological Virus Laboratory, Health Protection Agency, London, UK |
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Abstract: | Summary. The B-cell response against measles nucleoprotein (MeN) plays an important role in the control of measles infection. However,
the data on B cell epitopes of MeN are still limited. The objective of this study was to identify B cell epitopes in MeN using
monoclonal and polyclonal antibodies raised against recombinant yeast-expressed MeN (rMeN) as well as human sera from measles-positive
individuals. After immunization of mice, 15 monoclonal antibodies (mAbs) against rMeN were generated. The B cell epitopes
were localized using recombinant overlapping MeN fragments, PepScan analysis, and competitive ELISA. The epitopes of 14 mAbs
were mapped within the C-terminus of MeN between amino acids (aa) 419 and 525. Four mAbs recognized a linear epitope located
within a sequence of aa 440–448. Competitive ELISA revealed a cluster of conformational mAb epitopes. Cross-inhibition studies
with human sera demonstrated similar localization of B cell epitopes recognized by serum antibodies from naturally infected
individuals. Thus, the majority of B cell epitopes are located at the C-terminal domain of MeN. These findings provide new
data on the antigenic structure of MeN and are in agreement with recent experimental evidence indicating that the C-terminal
domain of MeN is well accessible on the surface of nucleocapsid-like structures. |
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