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Phage antibodies against an unstable hapten: Oxygen sensitive reduced flavin
Author(s) -
Bruggeman Yvonne E.,
Boogert Anja,
van Hoek Arie,
Jones Peter T.,
Winter Greg,
Schots Arjen,
Hilhorst Riet
Publication year - 1996
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(96)00544-3
Subject(s) - hapten , flavin group , antibody , chemistry , oxygen , flavin mononucleotide , biochemistry , phage display , riboflavin , microbiology and biotechnology , biology , enzyme , peptide , immunology , organic chemistry
It is difficult to raise antibodies against haptens and antigens that are unstable under the physiological conditions of the serum. Here we have used a phage antibody library to isolate antibody fragments against oxygen sensitive reduced flavin, by selection of the phage under anaerobic and reducing conditions at pH 5 and a pre‐elution step with the oxidized flavin. The binding of the reduced hapten to one of the antibody fragments was characterised by time‐resolved polarised fluorescence, and shown to be highly specific for the reduced flavin.