Unchained Labs eBook - 14

Unleash your biologic * Protein characterization turned up to 11

In the example shown in Figure 3, lysozyme was formulated
in a buffer with either 100 mM NaCl or 400 mM NaCl. The
solution in 400 mM NaCl became visibly cloudy when stored
at 4 °C. The cloudy appearance was reversible; the solution
clarified upon warming or dialysis into a lower salt buffer.
As expected, lysozyme exhibited a negative kD value of -6.0
mL/g in the 400 mM NaCl solution, and a positive kD value
of 3.8 mL/g in the 100 mM NaCl solution. The B22 values (not
shown) show the same trends, and were well-aligned with
measurements made by others using different techniques.

Conclusion
Uncle combines three detection methods to enable more
stability measurements on a single platform. This provides
more flexibility by enabling combinations of measurements
that can be performed simultaneously or in tandem. The
benefits of the Uncle platform are not limited to protein
characterization. As Virginie Maynard-Rose, Ph.D., Director
of Cell Line Development at Aragen Bioscience explains,
"The low protein requirements of the Uncle system make
this platform a key driver in early clone selection in antibody
campaigns for manufacturability assessment."

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GENengnews.com

Figure 3.
Diffusion coefficient as a function of lysozyme concentration at two different
salt concentrations, and the calculated kD values for each condition. The
positive value in the presence of 100 mM NaCl indicates repulsive interactions
between protein molecules. The negative value in 400 mM NaCl points to
self-association, which is consistent with visibly observable changes in that
protein solution.
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