RESUMO
High-concentration antibody solutions (>100 mg/mL) present significant challenges for formulation and process development, including formulation attributes such as increased solution viscosity, and the propensity for self-association. An additional challenge comes from the adaptation of analytical methods designed for low-concentration formulations to the high-concentration regime. The oligomeric state is a good example: it is a quality attribute monitored during pharmaceutical development and is one that can be affected by dilution; a typical first step in the analysis of high-concentration solutions. The objective of this work was to develop a size-exclusion HPLC (SE-HPLC) method that would allow the injection of high-concentration antibody formulations without the need for dilution prior to injection and their analysis in a high-throughput manner that does not create a bottleneck for the execution of complex formulation development studies. It was found that changing the UV detection wavelength from 215 to 235 nm simplified sample preparation by allowing for an approximately fivefold increase in injection load while maintaining the signal within the linear range of detection. In addition, the chromatographic peak properties (i.e., peak symmetry, resolution, and sensitivity) were determined to be consistent when compared with analytical methods developed for formulations with lower antibody concentrations.
