Kinetic Analysis of Synthetic Analogues of Linear‐Epitope Peptides of Glycoprotein D of Herpes Simplex Virus Type 1 by Surface Plasmon Resonance

Abstract

The interaction between mAb A16 and glycoprotein D (gD) of herpes simplex virus type 1 was analyzed by studying the kinetics of binding with a surface-plasmon-resonance biosensor. mAb A1 6 belongs to group VII antibodies, which recognize residues 11 -19 of gD. In a previous study, three critical residues, Aspl3, Argl6 and Phel7, of this epitope were identified by screening a phage display library that contained a random 15-amino-acid insert with the antibody. The contribution to binding of these residues in the motif DXXRF was further analyzed by an amino-acid-replacement study of the epitope gD-(9-19)-peptide and of a gD-(9-19)-peptide mimotope, previously obtained by screening the phage display library. Amino acid residues of the motif were replaced by a neutral amino acid residue, an amino acid residue with opposite charge and a corresponding D-amino acid residue. Kinetic parameters of peptide analogues were determined with a surface plasmon-resonance biosensor. The kinetic parameters of the peptide analogues were compared with the kinetic parameters of the interaction between mAb A16 and the epitope gD-(9-19)-peptide. The minimal size of the gD epitope for mAb A16 was also determined in this study. The kinetic constants of the resulting gD-(ll-l7)-peptide were found to be similar to those of entire gD. The kinetic analysis precisely defined the epitope on gD for mAb A16 to residues 11 -17, identified Argl6 as an essential residue and suggested that Asp13 and Phel7 are mainly involved in stabilization of the secondary structure of the peptide.Keywords: herpes simplex virus 1 ; glycoprotein D; linear epitope ; kinetics ; surface plasmon resonance.Glycoprotein D (gD) of herpes simplex virus (HSV) type 1 is an essential component of the virion envelope for the entry of HSV into mammalian cells [I] and is one of the HSV proteins on which immunological and vaccine studies have focused. Immunization of mice with gD of HSV-1 results in high neutralizing-antibody titers against HSV-1, and these mice are protected against a lethal HSV challenge [ 2 ] . mAbs directed against different regions of gD of HSV-1 have been divided into groups according to their antigenic sites [3], mAbs which recognize amino acid residues 11 -19 in the N-terminus are designated as group VII mAbs [4, 51. mAb A16 is a group VII mAb that binds a synthetic peptide consisting of amino acid residues 9-19 of gD [6, 71. Immunization with synthetic peptides comprising this region of HSV gD protected mice against a lethal challenge with HSV [8, 91. Substitution studies showed that Pro14 and Argl6 are important for binding of mAb A16 [7]. Additional information on residues that interact with mAb A16 was obtained by screening a random peptide library displayed on a filamentous bacteriophage [lo] with the antibody.Peptide sequences obtained by screening such a library resemble or mimic the epitope and are designated as mimotopes. We identified one mimotope. Its smallest synthetic version able to inhibit binding of the gD-(9-19)-peptide to mAb A16 was a ...

Functional Region IV of Glycoprotein D from Herpes Simplex Virus Modulates Glycoprotein Binding to the Herpesvirus Entry Mediator

“…Rabbit anti-gB (R69) and anti-gC (R46) sera (9) were used in immunoperoxidase assays. Anti-gD MAb DL6 (antigenic group IIb), which recognizes a continuous epitope from residues 272 to 279 (8,16), and anti-gD MAb ID3 (group VII) (10,21), which recognizes a continuous epitope from residues 11 to 19 (4, 7), were used for immunoaffinity purification and for analysis of antigenic activity. Anti-gD MAbs HD1 (group Ia) (27,35), DL11 (group Ib) (5,27), VID (group IIIa) (28,36), ABD (group IIIb) (36), 45S (group IV) (37), DL2 (group VI) (5), D4 (group IX) (14), and AP7 (group XII) (3,25) recognize discontinuous epitopes and were used for analysis of antigenic structure.…”

Partial d -amino acid substitution: Improved enzymatic stability and preserved Ab recognition of a MUC2 epitope peptide

“…However, most of the available literature reports on peptides in which all L-amino acid residues were replaced by their D-enantiomers, leading to normal (in all-D-isomer peptides) or reversed (in retro-all-D-isomer peptides) amide linkage (11,12). Only a few studies have been published on immune recognition of linear oligopeptides partially substituted by D-amino acid residue(s) in the epitope sequence (13,14). Apart from our previous studies (15,16), to the best of our knowledge, no data are available on the Ab binding to epitope peptides containing D-amino acid residues that are not in the core but in the neighboring N-and͞or C-terminal flanking region.…”

Peptide mimics elicit antibody responses against the outer-membrane lipooligosaccharide of group B Neisseria meningitidis

“…MAb 9-2-L379 (150 nmol) was pre-incubated with various concentrations of SMYGSYN for up to 3 h at ambient temperature [22] and reacted with NmLOS3,7,9 im-mobilised to a biosensor cuvette [19]. Non-speci¢c interaction between the SMYGSYN peptide and the immobilised NmLOS3,7,9 was subtracted from the binding pro¢les of mAb 9-2-L379 using the Fastplot1 software (A¤nity Sensors).…”