Synthesis and comparison of antibody recognition of conjugates containing herpes simplex virus type 1 Glycoprotein D Epitope VII~1

摘要

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 antibodly binding could be altered by conjugation to macromolecules or by modification in the flanking regions.However,no systematic studied have been reported to describe the effect of different carriers macromolecules in epitope conjugates.To this end,the influence of carrier structure and topolgy on antibody recognition of attached epitope has been studied by comparing the antibody binding properties of a new set of conjugates with tetratuftin 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 ~9LKNleADPNRFRGKDL~22 (Nle~11]-9-22)representing an immunodominant B cell epitope of herpes simplex uirus type 1 glcyoprothein D(HSV-1)gD)was conjugated to polypeptidesthrough 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 implied by-amino group of the polypeptides and its use for the conjugation of epitope peptides possessing Cys at C-terminal position.We have performed bindign studies (direct and competitive ELISA)with monoclonal antibody (Mab)A16,recognizing the HSV gD-related epitope,[Nle~11-9-22,and conjugates containing indentical and uniformy oriented epitope peptide in multiple copies attached to five different macromolecules as carrier.Data suggest that the chemial nature of the caarrier and the degree of substitution have marked influence on the strength of antibody binding.