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Preparation of Monomeric Fab'‐Horseradish Peroxidase Conjugate Using Thiol Groups in the Hinge and Its Evaluation in Enzyme Immunoassay and Immunohistochemical Staining. a
Author(s) -
ISHIKAWA ElJI,
YOSHITAKE SHINJI,
IMAGAWA MASAYOSHI,
SUMIYOSHI AKINOBU
Publication year - 1983
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1111/j.1749-6632.1983.tb22191.x
Subject(s) - horseradish peroxidase , conjugate , immunoassay , immunohistochemistry , chemistry , peroxidase , staining , enzyme , thiol , chromatography , monomer , microbiology and biotechnology , biochemistry , antibody , organic chemistry , pathology , medicine , biology , immunology , mathematical analysis , mathematics , polymer
Horseradish peroxidase and Fab' were conjugated by using thiol groups in the hinge of Fab'. Maleimide or pyridyl disulfide groups were introduced into peroxidase by treatment with N-succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate or N-succinimidyl 3-(2-pyridyldithio)propionate and were allowed to react with thiol groups in the hinge of Fab'. The conjugates were obtained in high yields with a minimal polymerization and without impairing the activities of peroxidase and antibodies, and were superior to those prepared using amino groups of Fab' by the glutaraldehyde and periodate methods in performing sandwich enzyme immunoassay and immunohistochemical staining. The conjugate yield was higher in the maleimide method than in the pyridyl disulfide method.