Reactivity of Anti-peptide and Anti-poliovirus Type 3 Monoclonal Antibodies with Synthetic Peptides

Abstract

SUMMARYMonoclonal antibodies were prepared from mice immunized with an 18-residue synthetic peptide with an amino acid sequence from a major antigenic sequence involved in the neutralization of type 3 poliovirus. Approximately 250 hybridomas secreted antibodies that reacted with the peptide but not the virus, two antibodies reacted with the virus but not the peptide and no antibody reacted with both. Conversely 26 monoclonal antibodies prepared from mice immunized with type 3 poliovirus and known to be directed against the appropriate sequence on the virus, generally failed to react with the peptide. These results might be expected if only a small proportion of the free or coupled peptide molecules adopt molecular conformations which resemble that of the homologous antigenic site in the virus. Antibodies specific for other antigens occasionally reacted well with the synthetic peptides, indicating that antibodies may bind to peptides of inappropriate sequence. The identification of antigenic sites by the use of synthetic peptides therefore requires considerable caution.We have used indirect biological methods to identify antigenic sites involved in the neutralization of type 3 poliovirus Evans et al., 1983). Three sites have been identified, an immunodominant site at amino acids 89 to 100 on VP1, a site on VP2 and a complex site composed of residues from VP3 and VP1 . These findings are consistent with the published X-ray crystallographic structure of poliovirus (Hogle et al., 1985) and with studies using polyclonal sera ). Other workers have used different approaches to identify antigenic sites on type 1 poliovirus. These include cross-linking of monoclonal antibodies to the virus (Emini et al., 1982), the induction of neutralizing antibodies by synthetic peptides (Chow et al., 1985) and the binding of neutralizing monoclonal antibodies to a VPl-fl-lactamase fusion protein (Van der Werf et al., 1983). In addition the ability of monoclonal or polyclonal antibodies to bind to synthetic peptides in direct ELISA has been used as a criterion for the identification of important antigenic sites in poliovirus type 1 (Emini et al., 1983), hepatitis B virus (Neurath et al., 1984) and Epstein-Barr virus (Dillner et al., 1984) and has been suggested as a general method (Geyson et al., 1984). We have therefore examined the immunological properties of synthetic peptides whose sequences were based on the known immunodominant site of type 3 poliovirus. The findings suggest that the binding of antibodies to synthetic peptides in direct ELISA cannot be regarded as a universally valid approach, and that the results of such studies should be interpreted with caution.The peptides used in these studies were synthesized by the Fmoc-polyamide method (Cambridge Research Biochemicals, Cambridge, U.K. ; Brown et al., 1983) and their sequences are given in Table 1. Synthetic peptide S10a, which contains the amino acid sequence of the major antigenic site on VP 1 of the Sabin vaccine strain of type 3 poliovirus, has been shown to induce ty...

Characterization of monoclonal antibodies raised against a synthetic peptide capable of inducing a neutralizing response to human rhinovirus type 2

“…MAbs in the remaining two groups all recognized intact virus particles, judged by their capacity to precipitate them efficiently, but could be differentiated by their ability to neutralize (group 1) or not (group 2). The high proportion of MAbs found in this study that recognize both virus and peptide contrasts with a previous report by Ferguson et al (1986). This group raised approximately 250 MAbs to peptides representing an antigenic site on poliovirus type 3, but found that the vast majority were peptide-specific, and the two that bound to virus did not recognize immobilized peptide.…”

“…Crystallization studies of neutralizing anti-peptide MAbs with virus and peptides could provide valuable information on the requirements of peptide design necessary to achieve this objective, and such studies are underway with the group 1 MAbs. Anti-peptide MAbs may also prove useful in the evaluation of novel presentation systems for synthetic antigens, particularly during the manipulation of immunogenic sequences that are not recognized by anti-viral MAbs in isolation because these sequences normally contribute to conformational sites, as in the case of poliovirus peptides (Ferguson et al, 1986). In this context we have used the MAbs described here to compare the HRV2 sequence expressed at two locations on hepatitis B core antigen particles (Brown et al, 1991).…”

A protective anti-peptide antibody against the immunodominant site of the A24 Cruzeiro strain of foot-and-mouth disease virus and its reactivity with other subtype viruses containing the same minimum binding sequence

“…Comparison of the sequences comprising 149-154 of VPI of a number of A subtype viruses (Weddell et al, 1985;Beck & Strohmaier, 1987) revealed that this region was reasonably conserved within a region of hypervariability, and often included only a single amino acid change. Based on this and also the observation that a single mutational change within an epitope may also enhance binding and neutralization by an antibody (Ferguson et al, 1986;Barnett et al, 1995) it was decided to determine if C 1.1 could cross-react with other A type strains. Preliminary assessment using a panel of 12-mer peptides encompassing VP1 residues 147-158 representing at least 17 different A strains indicated that almost all of the naturally occurring mutations within the minimum binding sequence abrogated binding.…”

Monoclonal antibodies to a peptide of human rhinovirus type 2 with different specificities recognize the same minimum sequence

“…Enhanced neutralization of a mutant virus (VP2 N163Y) was also seen with the anti-viral MAbs 801 and 829. Increased neutralization resulting from a single amino acid substitution has only been reported once previously (Ferguson et al, 1986). An anti-peptide M A b was shown to neutralize a mutant of poliovirus with a single substitution in VP1 threefold more efficiently than the parent virus.…”