Use of HPLC as an Enabler of Process Analytical Technology in Process Chromatography.

Abstract

Online monitoring of product quality attributes using high resolution analytical tools is a prerequisite for implementation of process analytical technology (PAT) and thereby ensuring product quality and consistency. Online high-performance liquid chromatography (HPLC) has been established for real time monitoring of product quality attributes. However, requirement of liquid handling system capable of online sampling and fractionation and interfacing it with preparative scale chromatography appends to the cost and complexity of the design module of commercially available online HPLC. This paper proposes a cost-effective approach for using a traditional offline HPLC for online analysis using a 2 way/6 port valve to facilitate simultaneous automated sampling of product stream eluting from a process column and fractionation. No sample dilution is required in the proposed approach. The versatility of the proposed online configuration has been verified by demonstrating its use for two of the most common separations required during production of monoclonal antibody therapeutics: separation of aggregates and separation of charge variants. Process modeling has been performed to allow interpolation of HPLC data and facilitate pooling to achieve the desired purity (model predicted purity 99.1% vs achieved purity of 99.3% for removal of aggregates, model predicted main species yield of 62.4% vs achieved main species of 62.9% for pooling of charge variants). The study thus demonstrates that the proposed online HPLC configuration can be used for PAT applications in preparative chromatography.