Influence of Column Temperature on Reversed-Phase Chromatography of an Intact Antibody

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Vanquish Flex UHPLC System was used in combination with the MAbPac RP column for the reversed-phase analysis of intact bevacizumab. Column thermostatting strongly influences peak shape and recovery. For runs with column temperature set between 60 and 90 °C, peak width at half height is below 3 seconds for a 30 minutes long gradient. The largest peak area is obtained at 70 °C, and decreases steeply at higher and lower column temperatures. Carry-over decreases at higher temperature. At 70 °C the carry-over is estimated at 0.37%.