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Real-time Transition Analysis curve in column chromatography with SIMCA®

Transition Analysis (TA) is a method traditionally used to evaluate the condition and integrity of a chromatography column. However, this method generally requires conducting complex data post‑processing and calculations which are hard to automate and to run in real time during the manufacturing process. Thus, completing the TA offline leads to time-consuming efforts that may delay the subsequent manufacturing operations. Colleagues from Amgen Manufacturing Limited developed an automated method using SIMCA® and SIMCA®-online to compute the TA curve in real time, allowing them to monitor and verify the condition of their column chromatography steps with limited operator input and no impact on manufacturing rate.

Introduction

Column chromatography steps are among the main unit operations in the purification area of any biopharmaceutical manufacturing setup. To ensure consistent product quality, it is essential to regularly monitor performance for consistency, degradation, efficiency and integrity. There are multiple techniques that have been used to evaluate chromatographic performance. One traditional technique is a pulse injection integrity test, generally used to assess the efficiency of a newly packed column.1 This method requires conducting an additional activity at the column to generate the data needed to calculate the height equivalent of a theoretical plate (HETP) and Asymmetry factor (Af) based on the normal distribution of an elution peak.

Transition Analysis (TA) is another promising technique for evaluating column performance. TA consists of measuring and inspecting the response of a signal (eg, conductivity) at the column outlet to a step change caused by an input solution,2,3 which can be measured during normal column operations, avoiding the additional activity required for the pulse injection integrity test. When performing TA, a first derivative transition analysis curve (TA curve) is calculated on the chromatographic transition,1,4 leading to a bell-shaped curve. HETP and Af can also be computed based on this transition curve.

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