Integration of Asymmetrical Flow Field-Flow Fractionation (AF4) with online native mass spectrometry (MS) is an attractive concept that offers significant value for the analytical characterization of protein therapeutics. Its execution, however, is challenged by the incompatibility of mobile phase components and the difficulties of achieving sensitive MS detection without compromising the separation efficiency. Herein, we describe in detail the successful coupling of a commercial AF4 instrument...

In the biopharmaceutical industry, the sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) assay is often used to assess therapeutic critical quality attributes (CQAs). Traditional detection methods for SDS-CGE methods, such as ultraviolet (UV) absorbance and laser-induced fluorescence (LIF), are widely used but come with limitations. A native fluorescence detection (NFD) scheme was previously developed to enhance sensitivity and reduce gel matrix interference without requiring sample...

Cell lines that produce non-originator versions of the National Institute of Standards and Technology (NIST) monoclonal antibody reference material 8671 (NISTmAb) are invaluable to the biopharmaceutical industry because, unlike typical commercial cell lines, they can be used on a collaborative and noncompetitive basis for bioprocess development. NIST has generated NS0 clones, NISTCHO research-grade test material 10197 and reference material 8675 NISTCHO to fill this need. We set out to optimize...

Density gradient equilibrium analytical ultracentrifugation (DGE-AUC) was first introduced in 1957. The method saw significant use over the following decade. Since then, DGE-AUC has been used by polymer and genomic DNA fields. Emerging medicine has revived interest in the technique for characterization of cell and gene therapeutics. While several model-dependent approaches exist to determine density at any point along a density gradient at equilibrium, there is ample evidence in the vast density...

Well-characterized reference materials enable successful collaborations within the scientific community by establishing common reagents for benchmarking studies and reducing the barriers to sharing materials and information. Here, we report the development of NISTCHO, a recombinant Chinese hamster ovary cell line expressing a nonoriginator version of the NISTmAb IgG1. We evaluated candidate clonal cell lines in a fed-batch cell culture model to assess growth and productivity of the cell lines...