To achieve better humanization, repeated analysis, construction and test procedures are required. The design of grafted or resurfaced antibodies often involves an iterated approach where sequence designs are generated and tested in binding and/or functional assays. https://www.creative-biolabs.com/antibody-humanization.html

1. Design Map1
Methods2
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Grafting
Chain
Shuffling
ANTIBODY HUMANIZATION
Creative Biolabs
Antibody
Humanization
Services
Multiple humanization degrees
Integrated antibody affinity maturation
Proprietary approach to evaluate the
immunogenicity
Humanization to antibodies of other species
Sequence and structure analysis
Construction, expression, identification
and biopanning of humanized antibodies
Multiple-degree humanization
Affinity and immunogenicity verification
FEATURES:WHAT WE DO:
VL
gene
The design of grafted or resur-
faced antibodies often involves
an iterated approach where
sequence designs are generat-
ed and tested in binding and/or
functional assays. Repeated
analysis, construction and test
procedures are required to
achieve better humanization.
In CDR-, FR-, and SDR-grafting
methods, all or part of the
variable or framework regions
of both murine and human
antibody genes are amplified
to generate the recombinant
plasmids, which are finally
applied to produce humanized
antibodies in bacterial or mam-
malian systems.
Except for chain shuffling
method generating fully
humanized antibodies, grafting
methods can generate antibod-
ies of different humanization
degrees. The single or
combined replacements of
complementarity determining
regions, framework regions and
specificity determining residues
respectively of a rodent
antibody by the corresponding
regions of human antibody can
respectively generate the
CDR-grafted, CDR & FR grafted,
and SDR-grafted humanized
antibodies.
murine
human
VH
gene
CL
gene
CH
gene
chimeric
L gene
chimeric
H gene
L chain of murine
Ab against a
certain antigen
human H chain
phage library
G3P
H chain
human L chain
phage library
L chain
G3P
human H chain
with highest-affinity
to the antigen
panned human
H/L chains pair
with highest-affinity
to the antigen
fully
humanized
partially
humanized
Humanization Degrees3
murine
Analysis of
Donor Sequences
Construction of
Fv 3D Model
Choice of
Acceptor
Sequences
Identification of
Backmutations
Design of
Humanized
Sequences
Construction
of Humanized
Sequences
Test of
Humanized
Sequences
humanized human
No ？
Yes ？
SDRs
CDR1
CDR2
CDR3
FR1
FR2
FR3
FR4
Human CDR
residues
Human
FR1
Human
FR2
Human
FR3
Human
FR4
Murine Ab CDR-grafted
Humanized Ab
CDR & FR-
grafted
Humanized Ab
“Abbreviated”
CDR-grafted
Humanized Ab
SDR-grafted
Humanized Ab
Murine constant regions
Murine framework regions
Non-SDRs of murine CDRs
SDRs of murine CDRs
Human framework regions
Human CDR residues
In chain shuffling method,
the light chain of the rodent
antibody is replaced by light
chains in human antibody
libraries; the resulting hybrid
antibody library is screened by
panning against the certain
antigen. The heavy chain of the
selected hybrid antibody is
replaced by heavy chains of
human antibody library. Subse-
quent screening of this second-
ary chimeric library will produce
fully humanized antibodies.
in vivo