Phage antibodies against an unstable hapten: oxygen sensitive reduced flavin.
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.
Bruggeman
Y E
YE
Department of Biochemistry, Agricultural Unversity, Wageningen, The Netherlands.
Boogert
A
A
van Hoek
A
A
Jones
P T
PT
Winter
G
G
Schots
A
A
Hilhorst
R
R
D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000276:immunology; D017104:Bacteriophage M13 / Q000276:immunology*; D005415:Flavins / Q000276:immunology*; D006241:Haptens / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000276:immunology*; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010100:Oxygen
D000914:Antibodies, Viral; D005415:Flavins; D006241:Haptens; D007128:Immunoglobulin Fragments; C055966:immunoglobulin Fv; D010100:Oxygen