Abstract
Immunization with Human Papillomavirus (HPV) L1 virus-like particles or L2 capsid protein elicits neutralizing antibodies that mediate protection. A highthroughput and sensitive in vitro neutralization assay is therefore valuable for prophylactic HPV vaccine studies. Over several hours during infection of the genital tract, virions take on a distinct intermediate conformation, including a required furin cleavage of L2 at its N-terminus. This intermediate is an important target for neutralization by L2-specific antibody, but it is very transiently exposed during in vitro infection of most cell lines resulting in insensitive measurement for L2, but not L1-specific neutralizing antibodies. To model this intermediate, we describe a protocol to generate furin-cleaved HPV pseudovirions (fc-PsV), which deliver an encapsidated reporter plasmid to facilitate infectivity measurements. We also describe a protocol for use of fc-PsV in a high-throughput in vitro neutralization assay for the sensitive measurement of both L1 and L2-specific neutralizing antibodies.
Original language | English (US) |
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Pages (from-to) | 14B.5.1-14B.5.26 |
Journal | Current Protocols in Microbiology |
Volume | 2015 |
DOIs | |
State | Published - 2015 |
Keywords
- Antibody
- Furin-cleavage
- HPV neutralization assay
- Hpv l2
- Human papillomavirus
- Papillomavirus
ASJC Scopus subject areas
- Parasitology
- Microbiology
- Virology