Studies on immunoassays of peptide factors. IV. New synthesis of a gastrin/peroxidase conjugate.
We have shown that structurally well-defined homogeneous maleoyl-peptides are synthetically accessible. These anchor-modified peptide derivatives allow their selective covalent linkage to thiol-containing proteins via the maleimide-thiol procedure. Correspondingly mercaptosuccinylated horseradish peroxidase was reacted with N alpha-maleoyl-beta-alanyl-human-little gastrin-I-[2-17] to produce the gastrin/peroxidase conjugate in good yields at 1:1 stoichiometry. The conjugate exhibited full enzymatic activity and identical binding affinity to antigastrin antisera as the parent gastrin. This approach proved to be well suited for the preparation of enzyme labeled peptide factors as tracers for immunoassays.
Moroder
L
L
Mourier
G
G
Bovermann
G
G
Dufresne
M
M
Göhring
W
W
Gemeiner
M
M
Wünsch
E
E
D004797:Enzyme-Linked Immunosorbent Assay; D005755:Gastrins / Q000032:analysis* / Q000276:immunology; D006735:Horseradish Peroxidase / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007118:Immunoassay; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis; D010544:Peroxidases / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet; D013438:Sulfhydryl Compounds / Q000032:analysis
D005755:Gastrins; D007074:Immunoglobulin G; D013438:Sulfhydryl Compounds; C013550:gastrin 17; D006735:Horseradish Peroxidase; D010544:Peroxidases