High-throughput protein aggregate analysis of monoclonal antibodies using a novel dual-channel UHPLC instrument

Introduction

In the current scientific environment, there is an increased need for the rapid and robust high-throughput analysis of biotherapeutics, in particular monoclonal antibodies (mAbs). This is due to the rise in the use of mAbs for the treatment of diseases.1 As a result, standardized chromatographic methods and excellent reproducibility are essential for sample analysis in quality control (QC) laboratories. Although standard UHPLC systems can analyze samples simply and rapidly, there is typically only the option to use one stationary phase at any given time. As a result, analysts are limited regarding the number of injections that a chromatography system can perform, leading to less efficient sample analysis and additional costs. In the biomanufacturing pipeline, biopharmaceutical handling and storage can cause a number of unintentional size variants of the original product that are potentially harmful for human health. Size-exclusion chromatography (SEC) is considered a gold standard for monitoring the formation and level of mAb aggregates and fragments2 and is probably one of the most frequently performed analyses in QC laboratories.

In this study, the Vanquish Flex Duo system for Dual LC was used for the high-throughput analysis of a mAb. One hundred injections of bevacizumab were performed on two identical Thermo Scientific™ MAbPac™ SEC-1 size exclusion columns. SEC is commonly used for the highresolution separation of mAb aggregates and fragments.

A number of chromatographic parameters were determined—including analyte retention time, peak width, area, symmetry, and efficiency—highlighting the similar results between each of the channels of the Vanquish Flex Duo system for Dual LC and the low % RSD values obtained. These results were also obtained on a standard Vanquish Flex Quaternary UHPLC system for direct comparison.