Download Evaluation of Large Panels of Bispecific Antibodies

Evaluation of Large Panels of Bispecific Antibodies Using
Surface Plasmon Resonance
Lope A. Flórez1, Romel Bobby1, Guido Cappuccilli1, Maria Wendt1, Baptiste Tauzin1, John Munro2, Stephan Heyse1 and Hans Peter Fischer1
1Genedata
AG, Basel, Switzerland | 2Genedata Inc., Boston, MA, USA | Email: [email protected]
Summary
Bispecific antibodies (BsAbs) have traditionally been developed by first optimizing antibody fragments then reformatting to the final bispecific format at the engineering stage.
However, some effective BsAbs are rejected early in the process. Moreover, costly engineering efforts are wasted optimizing fragments with poor kinetic properties in the final
format. These issues are leading to a paradigm shift in BsAb development: screening in the final therapeutic format as early as possible (“in-format screening”). Here, we present an
end-to-end software solution to run in-format screens in high throughput using Surface Plasmon Resonance (SPR) and/or Bio-Layer Interferometry (BLI). The software keeps track
of clones and supports their construction, automatically calculates kinetic parameters starting from raw SPR/BLI data, and uses the calculated data and sensorgrams to select the
best hits for optimization. This workflow scales with the combinatorial complexity of the approach, ultimately reducing development times and providing better BsAb lead candidates.
In-Format Screening Using SPR/BLI Technology
Challenges of Developing Bispecific Antibodies
• Traditionally, novel BsAbs have been developed by running mono-selective optimization
campaigns and re-formatting the best hits into the final therapeutic format.
In-format screening is an effective way to identify constructs with enhanced potency and efficacy
in the final format. It proceeds in three stages:
• However, due to changes in the pharmacological profile of the re-formatted moiety, this
approach can yield a low productivity.
1. In-silico design of target molecules based on libraries of variable regions, linkers, constant
regions, and backbone vectors, followed by parallelized construct synthesis and highthroughput transient expression.
• The solution is to screeen in the final therapeutic format as early as possible, preferably in
high-throughput (hundreds to thousands of clones).
Ideal BsAbs
with desired kinetic profile in final
therapeutic format
2. Off-rate ranking of antigen dissociation behavior (off-rate) by Surface Plasmon Resonance
(SPR) or Bio-Layer Interferometry (BLI).
Hits from mono-specific campaigns
reformatted to final
therapeutic format
3. Iterative selection of best clones from off-rate ranking, followed by purification, full kinetic
profiling, and epitope determination.
Scaling up to hundreds or thousands of clones requires mastery of multiple logistic and data
analytics challenges. Genedata software empowers the organization to address these challenges.
A
High-throughput
design &
registration
Construct
Synthesis &
Cloning
Recombinant
Expression
B
Screening and
characterization
by SPR/BLI
C
Hit
Selection
VR-1
Lost potential:
False negatives not pursued
further due to poor behavior in
mono-specific campaigns
Waste:
Engineering efforts are spent
on optimizing false positives,
with poor properties in final
format
Low productivity:
Only a fraction of useful hits
found with this approach, in
spite of high cost
VR-2
Integrative Genedata Platform for Design, Screening, and Selection of BsAbs in the Final Therapeutic Format
B
Genedata Screener® automates the processing of SPR/BLI
data at every stage of the workflow - from off-rate ranking
to full kinetic characterization. It provides rich interactive
visualizations to remove quality artifacts and select the best
hits. Data from multiple instrument runs can be analyzed
jointly, even if acquired on different instruments or by
different groups.
The screenshot illustrates a side-by-side comparison of
BsAbs from a primary high-throughput off-rate ranking
assay with a secondary follow-up kinetic characterization to
determine affinity.
Instrument-agnostic
SPR/BLI data analysis
of in-format BsAbs in
Genedata Screener®
Genedata BiologicsTM captures clone
properties, assay and sequence data, as well
as progeny information and stores them in
an integrated database. Best hits can be
selected based on full information acquired
for each clone.
A
C
Genedata BiologicsTM
contains an advanced
in silico cloning and
lab logistics solution
for highly parallelized
molecule design and
expression, optimally
suited for BsAbs.
Conclusions
Screening as early as possible in the final therapeutic format can lead to significant
increases in the productivity of BsAb campaigns and the quality of the resulting hits.
Genedata enterprise software empowers the organization to run these campaigns in
high-throughput.
Benefits of the Genedata platform in a nutshell
Efficienct clone and isolate
tracking, even for complex
therapeutic formats
Highly automated SPR/BLI
data analysis independent of
instrument and throughput
Hit selection based on a
comprehensive assessment of
all assays, including SPR/BLI