Simultaneous analysis of monoclonal antibodies using a novel dual-channel UHPLC instrument and orthogonal chromatography

Introduction

Although standard UHPLC systems can analyze samples simply and rapidly, typically only one stationary phase can be used at any given time, although there are multiple features to monitor and each requires its own method.

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. The Vanquish Flex Duo system for Dual LC can address these problems using two simultaneous LC channels for the same sample.

In this study, we focused on the flexibility of the Vanquish Dual LC workflow. Orthogonal analyses were performed on the same sample at the same time. The two analyses use different elution solvents and different instrument methods, varying in data collection wavelengths, gradient, or isocratic conditions and different run times. A number of chromatographic parameters were determined—including analyte retention time, peak width, area, symmetry, and efficiency—highlighting the similar results between each LC channel of the Vanquish Flex Duo system for Dual LC and the low % RSD values obtained.

Experimental

Recommended consumables

• Deionized water, 18.2 MΩ∙cm resistivity
• Thermo Scientific™ MAbPac™ SEC-1 column, 5 μm, 300 Å, 4.0 × 300 mm (P/N 074693)
• Thermo Scientific™ MAbPac™ RP column, 4 μm, 2.1 × 100 mm (P/N 088647)
• Thermo Scientific™ MAbPac™ SCX-10 RS column, 5 μm, 2.1 × 50 mm (P/N 082675)
• Fisher Scientific™ sodium phosphate dibasic anhydrous (P/N 10440481)
• Fisher Scientific™ sodium phosphate monobasic anhydrous (P/N 10751135)
• Fisher Scientific™ sodium chloride (P/N 11964051)