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VH H
VH H
Camelid sdAb development
C H2
Fc
C H3
A novel approach to anƟbody discovery from GenScript
Single Domain Antibody (sdAb):
Camelids naturally produce antibodies composed only of heavy chains as compared to the conventional antibodies
(see below). The antigen-binding site of these unusual heavy chain antibodies is formed only by a single heavy chain
variable domain, designated as single domain antibodies (sdAbs). sdAbs combine the advantages of conventional
antibodies with important features of smaller molecule sdAb drugs. The unique and well-characterized properties of
sdAbs make them ideal building blocks for various antibody-based applications. GenScript offers a comprehensive set
of services for sdAb research and development.
Advantages of sdAbs as compared to conventional antibodies:
Ability to bind “hidden” epitopes especially in challenging targets
Ability to bind into cavities or active sites of protein targets
Pure monomer
Better tissue penetration, even penetrating blood-brain-barrier
Excellent stability
Easier to engineer bi-specific and tri-specific antibody
Advantages of Choosing GenScript:
VH
C H1
VL
VH H
VH H
CL
C H2
C H2
C H3
C H3
ConvenƟonal IgG
Camelid heavy chain anƟbody
Two heavy chains
Two heavy chains
Two light chains
No light chain
Twelve protein domains
Six protein domains
Full Fc
Full Fc
sdAb
A leading sdAb service provider with seasoned scientists and proprietary technologies
Owns exclusive patents on technologies related to sdAb optimization
Ability to engineer sdAbs capable of achieving extremely high affinity (KD= 4 pM)
A recognized service provider for sdAb-based therapeutics for major pharmaceutical companies
Routes for sdAb generation:
Direct sdAb isolation from naive sdAb phage display library (1010 diversity) containing camel, llama and alpaca
sdAb repertoire.
Generation of high affinity sdAbs by camelid immunization and subsequent antibody engineering work.
Approaches for llama immunization:
DNA immunization
Cell immunization
Lipoparticles immunization
Protein immunization
Peptides immunization
Combination immunization
Technical platforms:
Phage display platform
Yeast surface display platform
FASEBA screening platform
Epitope binning platform
Kinetic determination platform
Featured Services:
120
Development of a panel of new antibody drugs,
targeting a key cytokine in cell growth
100
Candidate 2
80
Candidate 3
% Cell Viability
Case 1:
An immunized sdAb phage display library was constructed
from PBMCs after animal immunization. A panel of sdAbs
with good expression, high stability and affinity were
isolated using GenScript’s patented technology-FASEBA
platform. Three independent sdAbs were identified based
on high-affinity binding to the antigen, and were evaluated
by a cytokine-induced cell-proliferation assay. As shown by
the data in the graph below, their potency of inhibitory
activity was determined to be indistinguishable from that of
a marketed traditional antibody drug against the same
cytokine.
Candidate 1
Marketed Antibody Drug
60
40
20
0
10 -4
10 -3
10 -2
10 -1
10 0
10 1
10 2
Ab Concentration, [μg/ml]
Inhibition of cells proliferation by marketed antibody drug and top three
sdAb candidates
25
Development of a high affinity sdAb, as a
valuable reagent
An sdAb phage display library was generated using PBMC
and spleen lymphocytes. sdAb swere isolated and
characterized from this library, and the lead sdAb exhibits
good specificity and extremely high affinity. The specificity
and high affinity were validated by Biacore. The small
dimension (15 KDa,4×2.5×3 nm) of sdAb and super high
affinity make it excellent probe in biosensor applications.
100 nM
20
Response RU
Case 2:
25 nM
15
6.25 nM
10
6.25 nM
repeat
5
1.56 nM
0.39 nM
0
0
500
1000
1500
2000
Time s
Sensorgram of affinity measurement of the lead sdAb
Kinetics of the high affinity sdAb
Case 3:
Isolate sdAb for supporting protein
crystallization
kd (1/s)
KD (M)
5.73E+05
3.92E-05
6.83E-11
1.6
Absorbance/450nm
sdAbs are versatile crystallization chaperone that has
been recognized in this field. sdAb has a highly
structured β-sheet core that provides a rigid scaffold for
stabilizing potential intra-crystal contacts. In this case,
a batch of sdAbs was isolated from GenScript’s
ready-to-use naïve libraries, which specifically
recognize activated GPCR complexes but not ligand or
inactivated GPCR. After two rounds of subtractive
panning, the phage binders were enriched up to 6,000
fold. The sdAbs were chosen for supporting GPCR
crystallization.
ka (1/Ms)
Activated GPCR
Inactivated GPCR
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Toll-Free: 1-877-436-7274
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0.8
0.4
0
1
2
3
4
5
6
7
8
9
10 11 12 13 14 15 16 17 18 19 20 21 22
Several binders were obtained to positively recognize activated
GPCR only
Tel: 1-732-885-9188
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Fax: 1-732-210-0262
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Email: [email protected]
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860 Centennial Ave., Piscataway, NJ 08854, USA
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