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PDBsum entry 3l7e
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Immune system
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PDB id
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3l7e
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References listed in PDB file
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Key reference
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Title
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Human framework adaptation of a mouse anti-Human il-13 antibody.
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Authors
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J.Fransson,
A.Teplyakov,
G.Raghunathan,
E.Chi,
W.Cordier,
T.Dinh,
Y.Feng,
J.Giles-Komar,
G.Gilliland,
B.Lollo,
T.J.Malia,
W.Nishioka,
G.Obmolova,
S.Zhao,
Y.Zhao,
R.V.Swanson,
J.C.Almagro.
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Ref.
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J Mol Biol, 2010,
398,
214-231.
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PubMed id
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Note: In the PDB file this reference is
annotated as "TO BE PUBLISHED". The citation details given above have
been manually determined.
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Abstract
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Humanization of a potent neutralizing mouse anti-human IL-13 antibody (m836)
using a method called human framework adaptation (HFA) is reported. HFA consists
of two steps: human framework selection (HFS) and specificity-determining
residue optimization (SDRO). The HFS step involved generation of a library of
m836 antigen binding sites combined with diverse human germline framework
regions (FRs), which were selected based on structural and sequence similarities
between mouse variable domains and a repertoire of human antibody germline
genes. SDRO consisted of diversifying specificity-determining residues and
selecting variants with improved affinity using phage display. HFS of m836
resulted in a 5-fold loss of affinity, whereas SDRO increased the affinity up to
100-fold compared to the HFS antibody. Crystal structures of Fabs in complex
with IL-13 were obtained for m836, the HFS variant chosen for SDRO, and one of
the highest-affinity SDRO variants. Analysis of the structures revealed that
major conformational changes in FR-H1 and FR-H3 occurred after FR replacement,
but none of them had an evident direct impact on residues in contact with IL-13.
Instead, subtle changes affected the V(L)/V(H) (variable-light
domain/variable-heavy domain) interface and were likely responsible for the
5-fold decreased affinity. After SDRO, increased affinity resulted mainly from
rearrangements in hydrogen-bonding pattern at the antibody/antigen interface.
Comparison with m836 putative germline genes suggested interesting analogies
between natural affinity maturation and the engineering process that led to the
potent HFA anti-human IL-13 antibody.
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