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5
APPLICATION NOTE
Static light scattering experiments
Though the EAP Panta was not yet optimized for SLS experiments, research has
demonstrated the importance of looking at the SLS deviation from linearity as a more
reliable method for predicting aggregation behavior at high concentrations
1
. Using
scattering intensity, it was possible to build a ranking system similar to previous SLS
deviation experiments, and compare those to data obtained from other systems. Figure 3
shows the results of these experiments for a bispecific Fc-fusion protein
1
.
130.00
80.00
30.00
-20.00
-70.00
-120.00
-170.00
Panta Plate reader Microcuvette
His pH 8.5 +
20nM NaCl
His pH 7.5 +
20nM NaCl
His pH 7.5 +
150nM NaCl
His pH 5.5 +
150nM NaCl
His pH 6.5 +
20nM NaCl
His pH 5.5 +
20nM NaCl
Bispecific Fc-fusion protein
SLS deviation from linearity (%)
Deviation
from
linearity
(%)
Figure 3: Systems that demonstrate
a negative deviation from linearity via
light scattering measurements are
considered better candidates than
those that exhibit positive deviation.
The evaluated rankings of a bispecific
Fc-fusion for each formulation are the
same for all three instruments
1
.
As research has shown, it is important to have an alternative method beyond k
D
for
evaluating self-interaction of candidate molecules
1
. SLS deviation from linearity is a
parameter that allows direct measurement of high-concentration samples in order
to use this evaluation method as a predictive model for high-concentration scale-up.
