Preservation of immunoreactivity in the photolabeling of the B72.3 human antibody.
A versatile photochemical method of labeling human antibodies is described. Labeling is achieved by photolyzing 4-azido-2,3,5,6-tetrafluoro-14C-methylbenzoate and the B72.3 human antibody in a buffer at physiological pH. The photochemically produced nitrene presumably inserts into bonds in the hydrophobic part of the antibody resulting in > 75% attachment of the photoprobe. An immunoassay of B72.3 with mucin (B72.3 antigen) reveals > 97% retention of immunoreactivity and suggests that photochemical labeling is a viable alternative for the conjugation of biomolecules.
Pandurangl
R S
RS
Department of Chemistry, University of Missouri, Columbia 65211, USA.
Karra
S R
SR
Kuntz
R R
RR
Volkert
W A
WA
D000345:Affinity Labels / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001565:Benzoates / Q000737:chemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D010777:Photochemistry; D010782:Photolysis
D000345:Affinity Labels; D000911:Antibodies, Monoclonal; D001565:Benzoates; C098120:methyl 4-azido-2,3,5,6-tetrafluorobenzoate