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Titolo:
A single H : CDR3 residue in the anti-digoxin antibody 26-10 modulates specificity for C16-substituted digoxin analogs
Autore:
Short, MK; Jeffrey, PD; Demirjian, A; Margolies, MN;
Indirizzi:
Massachusetts Gen Hosp, Dept Surg, Antibody Engn Lab, MGH E, Charlestown, MA 02129 USA Massachusetts Gen Hosp Charlestown MA USA 02129 Charlestown, MA 02129 USA Mem Sloan Kettering Canc Ctr, Dept Cellular Biochem & Biophys, New York, NY 10021 USA Mem Sloan Kettering Canc Ctr New York NY USA 10021 New York, NY 10021 USA
Titolo Testata:
PROTEIN ENGINEERING
fascicolo: 4, volume: 14, anno: 2001,
pagine: 287 - 296
SICI:
0269-2139(200104)14:4<287:ASH:CR>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
AMINO-ACID SUBSTITUTION; PHAGE DISPLAY; MONOCLONAL-ANTIBODY; HIGH-AFFINITY; LIBRARIES; MUTATIONS; MATURATION; SITE; COMPLEMENTARITY; RECOGNITION;
Keywords:
antibody specificity; bacteriophage display; digoxin; immunoglobulin fragments; protein structure;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Margolies, MN Massachusetts Gen Hosp, Dept Surg, Antibody Engn Lab, MGH E,149 13th St,Box 31, Charlestown, MA 02129 USA Massachusetts Gen Hosp 149 13th St,Box 31 Charlestown MA USA 02129
Citazione:
M.K. Short et al., "A single H : CDR3 residue in the anti-digoxin antibody 26-10 modulates specificity for C16-substituted digoxin analogs", PROTEIN ENG, 14(4), 2001, pp. 287-296

Abstract

We constructed Fab libraries of bacteriophage-displayed H:CDR3 mutants in the high-affinity anti-digoxin antibody 26-10 to determine structural constraints on affinity and specificity for digoxin. Libraries of mutant Fabs randomized at five or 10 contiguous positions were panned against digoxin andthree C16-substituted analogs, gitoxin (16-OH), 16-formylgitoxin and 16-acetylgitoxin. The sequence data from 83 different mutant Fabs showed highly restricted consensus patterns at positions H:100, 100a and 100b for bindingto digoxin; these residues contact digoxin in the 26-10:digoxin co-crystalstructure. Several mutant Fabs obtained following panning on digoxin-BSA showed increased affinity for digoxin compared with 26-10 and retained the wild-type (wt) Trp at position 100. Those Fabs selected following panning onC16-substituted analogs showed enhanced binding to the analogs. Replacement of H:Trp100 by Arg resulted in mutants that bound better to the analogs than to digoxin. This specificity change was unexpected, as C16 lies on the opposite side of digoxin from H:CDR3, Substitution of wt Trp by Arg appearsto alter specificity by allowing the hapten to shift toward H:CDR3, thereby providing room for C16 substituents in the region of H:CDR1.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 01/12/20 alle ore 22:39:24