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 |
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Authors: | A G Laman A O Shepelyakovskaya Kh M Boziev G V Savinov F A Brovko V A Nesmeyanov |
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Institution: | 1.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Pushchino Division,Russian Academy of Sciences,Pushchino, Moscow oblast,Russia;2.Moscow State University, Pushchino Division,Pushchino, Moscow oblast,Russia;3.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry,Russian Academy of Sciences,Moscow,Russia |
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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 μM 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 × 108 and 3.5 × 108 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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