Real‐time detection of mAb aggregates in an integrated downstream process

Abstract

The implementation of continuous processing in the biopharmaceutical industry is hindered by the scarcity of process analytical technologies (PAT). To monitor and control a continuous process, PAT tools will be crucial to measure real‐time product quality attributes such as protein aggregation. Miniaturizing these analytical techniques can increase measurement speed and enable faster decision‐making. A fluorescent dye (FD)‐based miniaturized sensor has previously been developed: a zigzag microchannel which mixes two streams under 30 s. Bis‐ANS and CCVJ, two established FDs, were employed in this micromixer to detect aggregation of the biopharmaceutical monoclonal antibody (mAb). Both FDs were able to robustly detect aggregation levels starting at 2.5%. However, the real‐time measurement provided by the microfluidic sensor still needs to be implemented and assessed in an integrated continuous downstream process. In this work, the micromixer is implemented in a lab‐scale integrated system for the purification of mAbs, established in an ÄKTA™ unit. A viral inactivation and two polishing steps were reproduced, sending a sample of the product pool after each phase directly to the microfluidic sensor for aggregate detection. An additional UV sensor was connected after the micromixer and an increase in its signal would indicate that aggregates were present in the sample. The at‐line miniaturized PAT tool provides a fast aggregation measurement, under 10 min, enabling better process understanding and control.

Portable Microfluidic Viscometer for Formulation Development and in Situ Quality Control of Protein and Antibody Solutions

“…In this context, microfluidic approaches are well suited given their low volume requirements, their exquisite flow control, as well as the cost benefit they offer. Several microfluidic devices have been proposed to characterize the biophysical properties of protein solutions, including for the measurement of viscosity of therapeutic proteins. − …”

Robust method for chromatogram shape analysis to improve early detection of performance drifts and adverse changes in process parameters during purification column operations

“…Several recent examples for the implementation of PAT have demonstrated the feasibility and benefits of investing in such approach. [2][3][4][5] While some of these examples have focused on implementing new in-line/at-line instruments and sensors, [6][7][8][9][10][11][12][13][14][15] others have focused on how to best utilize the available data to make predictions and real-time decisions using multivariate analysis. [8,[16][17][18][19][20][21][22][23][24] Biotechnology processes often consist of several unit operations fed by multiple input parameters.…”