Screening of multimodal ligands for the capture and polishing of antibody streams: a microfluidic approach for rapid optimization of chromatographic operating conditions

Inês Fernandes Pinto

Key information

Authors:

Inês Fernandes Pinto (Inês Fernandes Pinto)

Supervisors:

Ana Margarida Nunes da Mata Pires de Azevedo (Ana Margarida Nunes da Mata Pires de Azevedo); João Pedro Estrela Rodrigues Conde (João Pedro Estrela Rodrigues Conde); Maria Raquel Múrias dos Santos Aires Barros (Maria Raquel Murias dos Santos Aires Barros)

Published in

09/24/2018

Abstract

The selection and optimization of chromatography ligands for the purification of biopharmaceuticals is a demanding challenge for biotechnologists. In the particular case of monoclonal antibodies (mAbs), synthetic ligands comprising multiple types of interactions (multimodal) can provide process and economic advantages compared to protein-based affinity ligands. However, optimizing the operating window of these multimodal ligands requires the development of simple, effective and affordable highthroughput screening platforms. In this thesis, a novel microfluidics-based strategy to perform rapid screening of chromatography operating conditions in the context of the purification of mAbs from cell culture supernatants is presented. Increasingly integrated devices were developed, aiming at (i) performing a simultaneous optimization of multiple chromatography ligands; (ii) addressing a multiplexed detection of different target molecules in solution; (iii) integrating automated and sequential liquid flow in the device; and (iv) coupling miniaturized photosensors for on-chip signal read-out. Microfluidic optimization studies were performed using fluorescently labeled molecules (IgG, BSA) in plain buffer solutions or spiked in real cell culture supernatants, enabling the trending of important parameters (purity and recovery yield) in accordance to conventional laboratory scale assays. The main advantages provided by the miniaturized devices and their intrinsic differences relative to a traditional chromatographic operation are also explored and discussed.