The Agilent 1290 Infinity II 2D‑LC interfaced with a Flownamics Seg‑Flow system for control of upstream titer, product quality, and amino acid concentration

This application note demonstrates the use of the Agilent 1290 Infinity II 2D‑LC system in combination with the Flownamics Seg‑Flow system for automated bioreactor sampling as a UHPLC‑based process analytical technology (PAT) tool for upstream titer and product quality assessments of mAbs and fusion proteins.

This combination enables the determination of multiple upstream critical quality attributes (CQAs) using a single setup and saves time and costs during the development of biotherapeutics. 2D‑LC heart‑cutting combined with a flow split after the first dimension enables shorter run time, improved peak shape, and therefore faster results to control the process.

As examples, the following CQAs have been determined for a mAb: titer, size variants, and charge variants as well as amino acid concentration of the bioreactor cell culture. Other potential applications include but not limited to online RP‑HPLC, HIC, HILIC, affinity chromatography, reduced and nonreduced denaturing SEC, etc. to measure quality attributes of bioreactor samples.

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