Robustness assessment of cation-exchange chromatography with in-silico peak deconvolution in monoclonal antibody purification

Abstract

An in-silico tool with peak deconvolution for preparative chromatograms was developed for assessing the robustness for peak fluctuations depending on loaded sample quality. The peak positions of high-molecular-weight species (HMWS), target monoclonal antibody (Main), and low-molecular-weight species (LMWS) on a chromatogram of cation-exchange chromatography (CEX) were identified by second derivative processing. The deconvoluted peaks of HWMS, Main, and LMWS were calculated by a fitting algorithm at each identified peak position. The height of deconvolved peaks of each component was varied to mimic the change of loaded sample quality in the chromatogram, and the quality of the pooled fraction obtained from the mimic chromatogram was estimated. In addition, the relationship between the loaded sample quality and the pooled sample quality was shown to evaluate the robustness of chromatographic purification process for peak fluctuations depending on loaded sample quality. A major advantage of this robustness assessment method is to be performed using the results of only a single wet-lab experiment. We considered that it is a useful in-silico tool for rapidly assessing chromatography robustness and also product quality depending on upstream process required to achieve the target quality in the pooled fraction, at the early stage of biological pharmaceutical product development, especially monoclonal antibodies