Elsevier

Analytical Biochemistry

Volume 608, 1 November 2020, 113899
Analytical Biochemistry

In-line detection of monoclonal antibodies in the effluent of protein A chromatography with QCM sensor

https://doi.org/10.1016/j.ab.2020.113899Get rights and content
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Highlights

  • Specific and real-time detection of monoclonal antibodies in the breakthrough fluid from protein A chromatography column.

  • In-line quartz crystal microbalance sensor with in-place regeneration.

  • Low cost and modular quartz crystal microbalance sensor with lower detection limit compared to UV detection.

Abstract

A major drawback of the IgG capture step is the high cost of the protein A resin. For a better utilization of the resin, a continuous multi-column operation was recently proposed. In this method, accurate detection of leaking IgG is crucial to divert the breakthrough fluid from the waste to the next column and prolong the loading step without product loss. The detection of a breakthrough point as a change in UV absorption is based on a relatively small signal addition of IgGs to the bulk signal of host cell proteins. To achieve specificity, we used a quartz crystal microbalance and immobilized protein A as specific ligand on the sensor surface. We integrated the quartz crystal microbalance sensor in-line after the protein A column for real-time detection of IgGs in the breakthrough fluid. We show that this specific IgG detection in the breakthrough fluid can be more sensitive than with the UV detector. The use of the same product-specific ligand in the affinity column and in the sensor allows simultaneous in-line regeneration of column and sensor in a single step. Such a sensor could support cost-efficient load control during the entire continuous multi-column capture step in downstream processing.

Keywords

Quartz crystal microbalance
mAb
IgG
In-line sensor
Protein A
Continuous multi-column affinity chromatography

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