Photoreduction of monoclonal antibodies for conjugation and fragmentation.
The conjugation of enzymes, fluorescent or radioactive labels, cross-linkers and other moieties to antibodies is a commonly performed procedure in biochemical research. Using reduced disulphides, conjugation can be an inconvenient, multistep, time- and material-consuming process. We have developed a reduction technique based on UV irradiation, which lacks these drawbacks. Antibodies are irradiated in a sealed vial for a few minutes by a common laboratory UV source in the presence of stannous ions, following the depletion of atmospheric oxygen. The preparation may subsequently be conjugated with thiol-reactive probes such as maleimide derivatives, with no need for any prior purification or concentration. This simple, rapid and effective reduction and conjugation process results in a fully functional immunoglobulin conjugate that can be used for a variety of biochemical applications.
Ellison
D
D
St. Bartholomew's Hospital, London, England, UK.
Stalteri
M A
MA
Mather
S J
SJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000302:isolation & purification / Q000528:radiation effects*; D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000302:isolation & purification / Q000528:radiation effects; D051379:Mice; D010084:Oxidation-Reduction; D010777:Photochemistry; D011869:Radioligand Assay; D013438:Sulfhydryl Compounds / Q000032:analysis; D014466:Ultraviolet Rays
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D013438:Sulfhydryl Compounds