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  • Title: [A method for the preparation of adjuvant peptide mimetics of GMDP with the use of monoclonal antibodies and combinatorial libraries of peptides in the format of phage display].
    Author: Laman AG, Shepeliakovskaia AO, Boziev KhM, Savinov GV, Brovko FA, Nesmeianov VA.
    Journal: Bioorg Khim; 2010; 36(2):170-7. PubMed ID: 20531474.
    Abstract:
    A method for the preparation of peptide mimetics of GMDP which could exhibit adjuvant activity without the negative effects of GMDP is described. The search for peptides with GMDP-like adjuvant activity was performed using highly specific monoclonal antibodies against GMDP and combinatorial peptide libraries in the format of phage display. Various elution methods were used for the immunoaffinity enrichment of the libraries during the course of the preparation of highly active and specific peptides. A sole peptide (Arg-Val-Pro-Pro-Arg-Tyr-His-Ala-Lys-Ile-Ser-Pro-Met-Val-Asn, RN) was obtained by the elution of phage particles from the immunosorbent with a 1 -microM solution of the natural ligand (GMDP). Elution with a buffer with a low pH value (0.1 M glycine-HCl, pH 2.2) gave two other peptides: Ser-Gly-Arg-Val-Ala-Val-Ser-Pro-Asp-Ser-Pro-Leu-Phe-Tyr-Pro (SP) and Arg-Tyr-Gly-Gly-Ser-Val-Leu-Asn-Ile-Glu-Cys-Gln-Phe-Tyr-Gly (RG). Affinity constants for the RN and SP peptides proved to be 3.6 x 10(8) and 3.5 x 10(8) M(-1), respectively. The specificity of the interaction with the monoclonal antibodies was checked by the competitive displacement of the peptides from the antigen-antibody complex by GMDP. The RN peptide exhibited adjuvant activity similar to that of GMDP, but had no pyrogenic effect characteristic of GMDP. The described method could be used for the search for mimetics of biologically active low-molecular compounds.
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