A phage library-derived single-chain Fv fragment in complex with turkey egg-white lysozyme: characterization, crystallization and preliminary X-ray analysis.
Using phage-display, an anti-turkey egg-white lysozyme single-chain Fv fragment was selected from a naive light chain variable region repertoire in combination with a heavy chain variable region 'mini library' of anti-hen egg-white lysozyme single-domain binders (Ward et al., 1989). Whereas the selected VH domain alone binds somewhat better hen egg-white lysozyme than turkey egg-white lysozyme, but both with comparatively low affinity, the specificity of VH is converted by addition of the VL domain. Thus, the single-chain Fv fragment is more specific for turkey egg-white lysozyme, with markedly increased affinities towards both lysozymes. The complex of single-chain Fv with turkey lysozyme has been crystallized and characterized by preliminary X-ray analysis.
Küttner
G
G
Institut für Biochemie des Universitätsklinikums Charité der Humboldt-Universität zu Berlin, Abteilung Proteinstrukturforschung, Germany.
Keitel
T
T
Giessmann
E
E
Wessner
H
H
Scholz
C
C
Höhne
W
W
D000595:Amino Acid Sequence; D000818:Animals; D000915:Antibody Affinity; D000936:Antigen-Antibody Complex / Q000737:chemistry; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000235:genetics; D005260:Female; D007128:Immunoglobulin Fragments / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009113:Muramidase / Q000737:chemistry* / Q000276:immunology*; D010063:Ovum / Q000201:enzymology; D014422:Turkeys
D000936:Antigen-Antibody Complex; D007128:Immunoglobulin Fragments; C055966:immunoglobulin Fv; D004247:DNA; D009113:Muramidase