TY - JOUR
T1 - Epitope mapping of African swine fever virus (ASFV) structural protein, p54
AU - Petrovan, Vlad
AU - Murgia, Maria V.
AU - Wu, Ping
AU - Lowe, Andre D.
AU - Jia, Wei
AU - Rowland, Raymond R.R.
N1 - Funding Information:
Funded was provided by the Kansas National Bio and Agro-Defense Facility Transition Fund and the Kansas Bioscience Authority through a matching grant to Kansas State University’s Center of Excellence for Emerging and Zoonotic Animal Diseases, Award # 15-130. Monoclonal antibody preparation was funded by the U.S. Department of Homeland Security under IAA Award # HSHQDC-12-X-00122. We would like to thank Dr. Baker for the contribution of the pHUE expression vector.
Publisher Copyright:
© 2020
PY - 2020/4/2
Y1 - 2020/4/2
N2 - In the absence of a vaccine for African swine fever virus (ASFV), diagnostic tools are critical for early detection and implementation of control measures. Along with other immunogenic proteins, p54 is a good serological target for conducting ASF detection and surveillance. In this study, a panel of 12 mouse monoclonal antibodies (mAbs) was prepared against a baculovirus-expressed p54(60–178) polypeptide. Further screening showed that five mAbs were positive for reactivity against ASFV-infected cells and recombinant p54 proteins. Mapping studies using five polypeptides and 12 oligopeptides, showed that mAb #154-1 recognized a conserved polypeptide sequence, p54(65–75), and was placed into Group 1. Mabs #143-1 and #7 recognized a region covered by p54(93–113) and were placed into Group 2. Group 3 consisted of mAbs #101 and #117, which recognized p54(118–127). Sera from pigs infected with the low virulent OURT 88/3 strain recognized the same p54 region covered by the Group 3 mAbs. When tested in a neutralization format, only mAb #143-1 showed neutralization activity above background. Together, the results identify important antigenic and immunogenic regions located on p54, which provide new tools for improving ASFV diagnostics.
AB - In the absence of a vaccine for African swine fever virus (ASFV), diagnostic tools are critical for early detection and implementation of control measures. Along with other immunogenic proteins, p54 is a good serological target for conducting ASF detection and surveillance. In this study, a panel of 12 mouse monoclonal antibodies (mAbs) was prepared against a baculovirus-expressed p54(60–178) polypeptide. Further screening showed that five mAbs were positive for reactivity against ASFV-infected cells and recombinant p54 proteins. Mapping studies using five polypeptides and 12 oligopeptides, showed that mAb #154-1 recognized a conserved polypeptide sequence, p54(65–75), and was placed into Group 1. Mabs #143-1 and #7 recognized a region covered by p54(93–113) and were placed into Group 2. Group 3 consisted of mAbs #101 and #117, which recognized p54(118–127). Sera from pigs infected with the low virulent OURT 88/3 strain recognized the same p54 region covered by the Group 3 mAbs. When tested in a neutralization format, only mAb #143-1 showed neutralization activity above background. Together, the results identify important antigenic and immunogenic regions located on p54, which provide new tools for improving ASFV diagnostics.
KW - African swine fever virus
KW - Epitope mapping
KW - Monoclonal antibodies
KW - p54
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U2 - 10.1016/j.virusres.2020.197871
DO - 10.1016/j.virusres.2020.197871
M3 - Article
C2 - 32004574
AN - SCOPUS:85078860032
SN - 0168-1702
VL - 279
JO - Virus Research
JF - Virus Research
M1 - 197871
ER -