Synthesis and Comparison of Antibody Recognition of Conjugates Containing Herpes Simplex Virus Type 1 Glycoprotein D Epitope VII¹

Abstract

Synthetic oligopeptides comprising linear or continuous topographic B-cell epitope sequences of proteins might be considered as specific and small size antigens. It has been demonstrated that the strength and specificity of antibody binding could be altered by conjugation to macromolecules or by modification in the flanking regions. However, no systematic studies have been reported to describe the effect of different carrier macromolecules in epitope conjugates. To this end, the influence of carrier structure and topology on antibody recognition of attached epitope has been studied by comparing the antibody binding properties of a new set of conjugates with tetratuftsin analogue (H-[Thr-Lys-Pro-Lys-Gly](4)-NH(2), T20) sequential oligopeptide carrier (SOC(n)), branched chain polypeptide, poly[Lys(Ser(i)-DL-Ala(m))] (SAK), multiple antigenic peptide (MAP), and keyhole limpet hemocyanine (KLH). In these novel constructs, peptide (9)LKNleADPNRFRGKDL(22) ([Nle(11)]-9-22) representing an immunodominant B cell epitope of herpes simplex virus type 1 glycoprotein D (HSV-1 gD) was conjugated to polypeptides through a thioether or amide bond. Here we report on the preparation of sequential and polymeric polypeptides possessing chloroacetyl groups in multiple copies at the alpha- and/or epsilon-amino group of the polypeptides and its use for the conjugation of epitope peptides possessing Cys at C-terminal position. We have performed binding studies (direct and competitive ELISA) with monoclonal antibody (Mab) A16, recognizing the HSV gD-related epitope, [Nle(11)]-9-22, and conjugates containing identical and uniformly oriented epitope peptide in multiple copies attached to five different macromolecules as carrier. Data suggest that the chemical nature of the carrier and the degree of substitution have marked influence on the strength of antibody binding.

Design, structural, and immuno‐analytical properties of antigenic bioconjugates comprising a β‐amyloid‐plaque specific epitope

“…For example, in the case of a (9-22) epitope peptide of Herpes simples virus (HSV) glycoprotein D type 1 (gD-1) no significant influence of carriers was found. 18 In accord with this, NMR studies of SOC and OT20 conjugates containing (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22) Design and Properties of Antigenic Bioconjugates 95 epitope of HSV gD-1 showed that all four epitope-copies attached to the carriers had identical conformation and did not interact with each other or with the carrier. It was also determined that in the case of OT20-conjugate the C-terminus attachment site of the epitope was more flexible than in the case of SOC conjugate.…”

“…The chloroacetylated MAP construct (ClAc-K(ClAc)-K(ClAc-K(ClAc))RRbA-NH 2 , 11) was built up also on MBHA resin, with a lower coupling capacity (0.67 mmol/g), while the SOC derivative (Ac-[K(ClAc)AibG] 4 -OH, 12) was assembled on PAM resin (0.72 mmol/g). 18 A new chloroacetylated oligotuftsin derivative analogue, elongated at the N-terminus with a promiscuous helper T-cell epitope derived from a hepatitis B surface antigen (Ac-FFLLTRILTIPQSLD-[TKPK(ClAc)G] 4 -NH 2 , 10), was synthesized on MBHA resin (0.67 mmol/g capacity). The following side chain protected N a -Boc-amino acid derivatives were Ac-C-Ab4-10 Ac-CFRHDSGY-NH 2 6…”

Strategies and Limitations in Dendrimeric Immunogen Synthesis. The Influenza Virus M2e Epitope as a Case Study

“…One particularly successful approach has been multimerization, either by simple polymerization (7) or by chemically better defined strategies such as conjugation to sequential oligopeptide carriers (8,9), display on lysine scaffolds (10,11), or, last but not least, incorporation into dendrimeric systems such as the multiple antigenic peptides (MAPs) pioneered by Tam in the 1980s (12). MAPs are based on a small immunologically inert core of radially branching lysine dendrites onto which a number of peptide epitopes are attached.…”

Artificial Carriers: A Strategy for Constructing Antigenic/Immunogenic Conjugates

“…A modified peptide LKNleADPNRFRGKDL ([Nle 11 ]-9-22) representing an immunodominant B cell epitope of HSV-1gD [67] was conjugated to SAK through a thioester bond. Binding studies (direct and competitive ELISA) were performed with mAb A16, recognizing the HSVgD-related epitope and the SAK conjugate and the results were compared with other conjugates of different artificial carriers [68].…”