Fully synthetic immunogens. Part III. Synthesis of hinge-peptide/gastrin conjugates and their immunological properties.
As core molecule for the multiple attachment of antigenic peptides we have selected the human IgG1 hinge fragment 225-232/225'-232'. Two types of conjugates of this double-chain bis-cystinyl hinge-peptide were prepared i) by linking its C-termini to [NIe15]-human-little-gastrin-[2,17] and ii) by elongating the resulting hinge-peptide/[NIe15]-little-gastrin-[2-17] conjugate at the two N-termini with the human big-gastrin sequence 1-14 to produce the big-gastrin-[1-14]/hinge-peptide/little-gastrin-[2-17] conjugate. For the synthesis of these peptide structures both the route via the preformed double-chain bis-cystinyl peptide and the route via suitably protected monomeric bis-cysteinyl peptides were used. For the latter approach advantage was taken of the previous observation about the preferred oxidation of the bis-cysteinyl hinge-peptide 225-232 to the dimer in parallel alignment. Both synthetic routes led to identical products. Immunization experiments in guinea pigs with the synthetic hybrids led to surprisingly strong immune responses with anti-little-gastrin antibody titers comparable to those induced by the iso-1-cytochrome c/little-gastrin-[2-17] conjugate as carrier-hapten system. These findings show that the two gastrin constructs are fully competent immunogens. Additionally, the gastrin receptor-like specificity of the antibodies indicates that both the synthetic hybrids and the cytochrome c conjugate allow for expression of a little-gastrin-specific conformational epitope similar to the bioactive structure of this hormone. The usefulness of such synthetic hybrids is further confirmed by the observation that the bivalent immunogen, containing both the little-gastrin 2-17 and the big-gastrin 1-14 sequence, is capable of inducing an immune response against both antigenic sequences, although with different efficiency. These results fully confirm our expectations.
Wünsch
E
E
Max Planck Institute of Biochemistry, Department of Peptide Chemistry, Martinsried, FRG.
Moroder
L
L
Hübener
G
G
Musiol
H J
HJ
Von Grünigen
R
R
Göhring
W
W
Scharf
R
R
Schneider
C H
CH
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005755:Gastrins / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology*; D006168:Guinea Pigs; D006241:Haptens / Q000138:chemical synthesis / Q000276:immunology; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology; D011487:Protein Conformation; D014614:Vaccines, Synthetic / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D005755:Gastrins; D006241:Haptens; C068551:IgG hinge protein-gastrin 17 (2-17); D007074:Immunoglobulin G; D010446:Peptide Fragments; D014614:Vaccines, Synthetic; C068550:gastrin 34 (1-14)-IgG hinge protein-gastrin 17 (2-17)