TECHNICAL NOTE
5
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to patients, and potentially requires increased infusion volumes to dilute the sample.
Additionally, propensity to self-interact increases the likelihood of aggregates forming
during production and storage of a biologic therapeutic. Optimizing buffer formulation to
reduce the self-interaction propensity of a candidate is a key consideration of the biologic
development workflow.
Candidate selection and storage buffer optimization are done in the early stages of
biologic development, and the k
D
of a given formulation is an important parameter when
considering the best options for further development and scale-up processes. With the
Prometheus Panta, it is easy to do such measurements, as well as obtain other important
candidate ranking parameters such as T
m
, T
on
, r
H
, and turbidity monitoring3.
Materials and Methods
Comparison of NISTmAb and Lysozyme concentration impact on radius
NISTmAb was diluted in 25 mM Histidine pH 6.0 (Figure 1a&b, teal). Lysozyme was diluted
in 25 mM Histidine pH 5 buffer supplemented with 130 mM NaCl (Figure 1a, purple).
Five replicates of each sample at each concentration were prepared in high-sensitivity
capillaries (NanoTemper Technologies, GmbH). A high-sensitivity size analysis experiment
was run with Panta.Control so ware v1.0 and analyzed with Panta.Analysis v1.2.
Self-interaction analysis with NISTmAb
NISTmAb was diluted in both 25 mM Histidine pH 6.0 (Figure 2, purple) and 25 mM
Histidine pH 6.0 + 150 mM NaCl (Figure 2, teal). Each sample was prepared in triplicate in
high sensitivity capillaries (NanoTemper Technologies GmbH). Data was acquired at 25
o
C
in size analysis high sensitivity mode with ten acquisitions per sample (i.e. 30 data points
per concentration). Data analyzed with Panta.Analysis v1.1.
