Application of anti‐peptide antibodies to the assignment of the inter‐chain disulphide bond in tetanus toxin

Abstract
Cysteine-containing peptides corresponding to the putative hinge region connecting the heavy and light polypeptide chains of tetanus toxin were synthesized utilising a solid phase with an acid hyper-labile linkage agent. Both the single-chain cysteine peptides, as well as a disulphide-bonded double-chain peptide, obtained by selective iodine-oxidation of S-trityl and S-acetamidomethyl protected peptides, were conjugated to carrier proteins for the purpose of immunisation and immunoassay. Comparison of the immunochemical specificity of mouse antibodies raised against these constructs, as well as antibodies against tetanus toxoid, permitted the assignment of the location of the inter-chain disulphide bond of tetanus toxin.