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TCR Engineering and Characterization Service
TCR Engineering and Characterization
Service
Substantial Affinity Improvement
Universal Assay Format for High-throughput Functional Screening
Integrated solution for TCR
TCR library construction,
Effective affinity matura-
tion platform to substan-
tially improve affinity ( ~
1800-fold)
phage panning and
screening
Affinity Maturation
~ 50 TCR leads
Universal assay platform
to allow a high-through-
put for in vitro functional
evaluation and specificity
screening
Functional evaluation and
specificity screening
Functional
Characterization
~ 20TCR leads
Superior assay sensitivity
in detection of antigen
independent T cell
In vivo efficacy validation
In Vivo
Evaluation
activation to mitigate the
risk of off-target toxicity
2
TCR leads
Affinity Maturation
TCR design and expression validation in mammalian and prokaryotic platform
Binding validation of TCR formats in phage system
Multi-prong approaches of TCR affinity maturation in phage display platform
Characterization of affinity matured TCR variants expressed in mammalian cells
Functional Evaluation
Reporter cell-based assay (RGA) based on TCR-knockout Jurkat cells
Primary T cell based cytokine release
Primary T cell based cytotoxicity
Functional evaluation of TCRxCD3 bispecific by RGA
In Vivo Assay
Animal model development and in vivo efficacy evaluation
Pharmacokinetics for TCRxCD3 bispecific and in vivo persistence for TCR-T cells
In vivo safety & early toxicity evaluation
ProBioCDMO.com
+1-732-885-9188
Accelerating Ideas. Delivering Impact.
Case Study: TCR Affinity Maturation(NY-ESO-1)
1866 fold of affinity improvement was achieved
comparing with parental (wt) TCR
Workflow
scTCR format validation of
NY-ESO-1 with phage
supernatant by ELISA
Ligand
Analyte
WT TCR
PC TCR
dsTCR1
dsTCR2
dsTCR3
dsTCR4
dsTCR5
ka (1/Ms)
9.06E+03
1.67E+05
7.52E+04
9.94E+04
1.03E+05
1.05E+05
7.54E+04
kd (1/s)
5.93E-03
3.39E-04
5.88E-05
4.37E-05
3.62E-05
8.43E-05
5.15E-05
KD (M)
6.55E-07
2.03E-09
7.82E-10
4.39E-10
3.51E-10
8.04E-10
6.83E-10
NY-ESO-1
NY-ESO-1
NY-ESO-1
NY-ESO-1
NY-ESO-1
NY-ESO-1
NY-ESO-1
9 consecutive NNK libraries
constructed, followed by
panning & screening
Total 51 and 32 unique
variants identified for TCR α
and β chain after two rounds
of panning/screening
Affinity matured TCRs specifically bind to relevant
HLA-A*02:01-NY-ESO-1 but not irrelevant
7 combinatorial variants
selected for recombinant
expression & validation
ELISA binding to HLA-A*02:01-GP100
ELISA binding to HLA-A*02:01-NY-ESO-1
4 lead candidates
selected for further
characterization
Case Study: Functional validation of TCRxCD3 bispecific
Functional evaluation TCRxCD3 bispecific by RGA line has been validated with formats of antigens as
pMHC, relevant tumor cell, or peptide loaded T2 cell
Activation of RGA line with tumor cell
Tumor Cell Based T Cell Activation Assay
Tumor Cell Based T Cell Activation Assay
Cell line
HLA-A*0201 GP100
SK-MEL-5
+
+
-
+
-
A375
SK-MEL-28
+
Activation of RGA line with plate
coated pMHC
Activation of RGA line with peptide
loaded T2 cell
T cell Activation by Tebentafusp (100nM)
pMHC Based T Cell Activation Assay
T2 Cell Based T Cell Activation Assay