Linear Multiepitope (Glyco)peptides for Type-Specific Serology of Herpes Simplex Virus (HSV) Infections

Abstract

Detection of type-specific antibodies is an important and essential part of accurate diagnosis, even in silent carriers of herpes simplex virus (HSV)-1 (oral) and HSV-2 (genital) infections. Serologic assays that identify HSV-1 and HSV-2 type-specific antibodies have been commercially available for more than a decade but often face problems related to cross-reactivity and similar issues. Attempts to identify type-specific peptide epitopes for use in serology for both HSV-1 and HSV-2 have been limited. We recently demonstrated epitope mapping of envelope glycoprotein G2 and identified a type-specific glycopeptide epitope that broadly recognized HSV-2 infected individuals. In the present work we have performed a comprehensive glycopeptide synthesis and microarray epitope mapping of 14 envelope proteins from HSV-1 and HSV-2, namely, gB, gC, gD, gE, gG, gH, and gI, using sera from HSV-1- and HSV-2-infected individuals and control sera. Several unique type-specific peptide epitopes with high sensitivity were identified and synthesized as one large linear multiepitope sequence using microwave-assisted solid-phase (glyco)peptide synthesis. Microarray validation with clinically defined HSV and Varicella Zoster (VZV) sera confirmed excellent cumulative specificities and sensitivities.

A general approach for identifying protein epitopes targeted by antibody repertoires using whole proteomes

“…Also, this epitope has been validated as an HSV2-specific diagnostic [44,45]. The HSV1-specific epitope was also similar to the previously reported epitope DHTPPMPSIGLE [18]. Interestingly, the two HSV-specific epitopes terminated in an identical 7-mer sequence EGAGDGE (PMPSIGLEEEEEE EGAGDGE and GGPEEF EGAGDGE ) [42].…”

“…Notably, 11 of these epitopes were bound by all HSV2 specimens. K-TOPE identified a glycoprotein C epitope PRTTPTPPQ with 83% prevalence which was contained in a previously identified epitope RNASA PRTTPTPPQ PRKATK [18]. In contrast to the numerous HSV2-specific epitopes, only 4 HSV1-specific epitopes were identified, and the highest prevalence achieved was only 40% (Table 5).…”

Epitope-mapping of the glycoprotein from Crimean-Congo hemorrhagic fever virus using a microarray approach

“…37 clinically validated sera from individuals infected with other viral diseases such as TBE, (n = 9), EBV, (n = 10) [ 16 ] and HSV, (n = 9) [ 15 , 19 ] and VZV, (n = 9) [ 19 ] were included in the study.…”

A novel monoclonal antibody targeting carboxymethyllysine, an advanced glycation end product in atherosclerosis and pancreatic cancer

“…Peptides were synthesized as earlier described by Risinger et al . [ 28 ] on tip-supported beads (1.4 μmol scale), using automated 9-fluorenylmethoxycarbonyl (Fmoc) SPPS on a SyroI (Biotage, Uppsala, Sweden) with the software SyroXP. Two equivalents of Fmoc-L-CML(oBut)(Boc)-OH was used together with four equivalents of 20 standard Fmoc-protected amino acid building blocks and Fmoc-O2Oc-OH spacer and coupled onto TentaGel S RAM resin (0.24 mmol/g loading density) using standard SPPS solvents.…”

“…Microarray printing was performed as earlier described [ 28 – 31 ] on hydrogel coated Nexterion ® Slides H, NHS using a MicroGrid array printer (60 nL/deposit, quilled pins, 250 μm pitch, BioRobotics, Genomics Solutions, Cambridgeshire, UK) with TAS Application suite software. In brief, glycated peptides and proteins were dissolved in printing buffer (17 mM NaH 2 PO 4 ×H 2 O, 133 mM Na 2 HPO 4 ×2 H 2 O, 0.03% NaN 3 , pH 8.5) to a final concentration of 700 μM for peptides and 200 μM for proteins to ensure that the microarray spots were saturated with the printed targets.…”