MaE11

Antibody Name: MaE11
References: Presta, L.G., Lahr, S.J., Shields, R.L., Porter, J.P., Gorman, C.M. Fendly, B.M. and Jardieu, P.M. "Humanization of an antibody directed against IgE" J. Immunol. (1993) 151: 2623-2632.
Donor Antibody: mouse
Acceptor Antibody: Human IgG1 | k
Antigen: Human IgE (free, not bound to receptor) see also C21.
Laboratory: Genentech, CA
Design: Consensus sequences of most abundant human subgroups (VH SGIII | VL SGI). Model mouse variable regions plus humanised versions. Made 22 versions testing buried residues, unusual murine sequence and packing residues as in Padlan, E.A. "A possible procedure for reducing the immunogenicity of antibody variable domains while preserving their ligand-binding properties" Mol. Immunol. (1991) 28:489-498. Analysed F(ab)s first and then made full-length mAbs.
Frameworks: VH SGIII | VL kI consensus sequences from Kabat Ed. V (1991). Insertion just before CDR-H1.
CDRs: All 6 VH and VL, Kabat Definition except that some CDR residues were kept human (H2 - S57T L63V N65G L1 - K24R L2 - G55E)
Backmutations: VH A24V F27Y T28S F29I V37I F67I L78F
VL M4L
Binding: Best binder F(ab)-12 inhibits IgE binding 30% as well as mouse mAb. Note that verneered version F(ab)-10 had different affinity when expressed as whole IgG.
Expression: F(ab) fragments in E.coli and whole mAbs in adenovirus-transformed human embryonic kidney cell line 293.
Comment: Consensus sequences "eliminates possible idiosyncracies present in any individual FR". Tried using Chothia definition for CDR-H2, but needed to change some residues in region 60-65 back to mouse. CDR-H1 includes both Chothia and Kabat definitions.
Clinical Indication: IgE mediated allergies

explanation of symbols and abbreviations

José Saldanha © 1997-8. Birkbeck College, London WC1E 7HX.