Serological Analysis of Herpes B Virus at Individual Epitope Resolution: From Two-Dimensional Peptide Arrays to Multiplex Bead Flow Assays

Macacine herpesvirus
or B Virus (BV) is a zoonotic agent that leads
to high mortality rates in humans if transmitted and untreated. Here,
BV is used as a test case to establish a two-step procedure for developing
high throughput serological assays based on synthetic peptides. In
step 1, peptide microarray analysis of 42 monkey sera (30 of them
tested BV positive by ELISA) revealed 1148 responses against 369 different
peptides. The latter could be grouped into 142 different antibody
target regions (ATRs) in six different glycoproteins (gB, gC, gD,
gG, gH, and gL) of BV. The high number of newly detected ATRs was
made possible inter alia by a new preanalytical protocol that reduced
unspecific binding of serum components to the cellulose-based matrix
of the microarray. In step 2, soluble peptides corresponding to eight
ATRs of particularly high antigenicity were synthesized and coupled
to fluorescently labeled beads, which were subsequently employed in
immunochemical bead flow assays. Their outcome mirrored the ELISA
results used as reference. Hence, convenient, fast, and economical
screening of arbitrarily large macaque colonies for BV infection is
now possible. The study demonstrates that a technology platform switch
from two-dimensional high-resolution peptide arrays used for epitope
discovery to a readily available bead array platform for serology
applications is feasible.