Photo-activated affinity-site cross-linking of antibodies using tryptophan containing peptides.
Affinity-based conjugation methods for antibodies can produce defined and reproducible conjugates. This requires that the target antibody has an affinity site for the ligand and that the ligand has a reactive site. These requirements are critical for the conjugation of antibodies designed for diagnostic and therapeutic application. Our laboratory has discovered a novel affinity of antibodies for the amino acid tryptophan using an azido derivative of tryptophan. Here we show that tryptophan without the azido group can be photo-cross-linked to antibodies. Biotinylated tryptophan peptides are photolysed into monoclonal and polyclonal antibodies and such biotinylated antibodies are used in avidin-based ELISA. With the simple and gentle tryptophan-affinity photo-conjugation of peptides, antibodies can be conjugated with peptides to enhance their potency and expand their targeting range.
Russ
Mike
M
InNexus Biotechnology, ImmPheron Laboratories, Inc. UK Coldstream Research Campus, 1501 Bull Lea Road, Suite 105, Lexington, KY 40511, USA.
Lou
Dingyuan
D
Kohler
Heinz
H
D000818:Animals; D000915:Antibody Affinity; D001666:Binding Sites, Antibody / Q000502:physiology*; D019921:Biotinylation / Q000379:methods; D003432:Cross-Linking Reagents / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008027:Light; D051379:Mice; D016853:Muromonab-CD3 / Q000378:metabolism*; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D010782:Photolysis; D014364:Tryptophan / Q000276:immunology* / Q000378:metabolism
D003432:Cross-Linking Reagents; D016853:Muromonab-CD3; D010455:Peptides; D014364:Tryptophan