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Microfluidic Cartridge for Bead-Based Affinity Assays.

Inês F Pinto¹, Veronique Chotteau², Aman Russom^3,4

¹KTH Royal Institute of Technology, Division of Nanobiotechnology, Department of Protein Science, Science for Life Laboratory, Solna, Sweden.

Methods in Molecular Biology (Clifton, N.J.)

|May 16, 2024

Summary

This summary is machine-generated.

A novel microfluidic cartridge enables rapid, at-line quantification of key proteins and metabolites in biopharmaceutical manufacturing. This Process Analytical Technology (PAT) tool offers a cost-effective solution for optimizing bioprocesses and ensuring product quality.

Keywords:

Biopharmaceuticals Colorimetry Immunoassays Microfluidics Monoclonal antibodies Multiplexing Process analytical technology

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Area of Science:

Biotechnology
Analytical Chemistry
Process Engineering

Background:

The biopharmaceutical industry faces intense competition, necessitating efficient process optimization to reduce time and costs.
Mammalian cell culture processes for monoclonal antibody (mAbs) production are complex and difficult to optimize.
Existing analytical methods lack integration for real-time monitoring within bioreactors.

Purpose of the Study:

To develop and demonstrate a microfluidic cartridge for at-line protein quantification in bioprocess monitoring.
To address the unmet need for rapid, integrated analytical tools in Process Analytical Technology (PAT).
To showcase the potential of microfluidics for sensitive and efficient bioprocess analysis.

Main Methods:

Fabrication and handling of a microfluidic cartridge for bead-based affinity assays.
Geometrical multiplexed immunodetection of specific protein analytes directly from bioreactor samples.
Quantification of Chinese hamster ovary (CHO) host cell proteins (HCP), IgG, and lactate dehydrogenase (LDH).

Main Results:

The microfluidic device achieved protein quantification within 2.5 hours with minimal hands-on time.
Demonstrated limits of detection in the low ng/mL range for HCP, IgG, and LDH.
Exhibited negligible matrix interference and no cross-reactivity between on-chip immunoassays.

Conclusions:

The developed microfluidic assay holds significant potential for Process Analytical Technology (PAT) in biopharmaceutical manufacturing.
This miniaturized analytical method offers reduced cost and complexity compared to current state-of-the-art instruments.
Enables systematic monitoring and quantification of critical analytes for improved bioprocess control and optimization.