This document describes the analysis of trypsin-digested monoclonal antibody and bovine serum albumin by UHPLC using Agilent ZORBAX RRHD 300SB-C18 1.8 µm reversed-phase media. The media demonstrates robustness with different acidic eluents, temperatures, and flow rates, and exhibits good reproducibility.
This document presents an application note detailing the use of the Agilent ZORBAX RRHD 300SB-C18 1.8 µm reversed-phase media for analyzing trypsin-digested monoclonal antibodies (MAb) and bovine serum albumin (BSA). The study highlights the column's ability to resolve protein digests efficiently using UHPLC systems.
The note explores various analytical conditions, including different digestion times, temperatures (up to 90 °C), flow rates, and acidic eluents such as trifluoroacetic acid (TFA) and formic acid. It demonstrates that the column provides good reproducibility and stability, particularly when using formic acid for UHPLC-MS applications, while maintaining peak shape and efficiency.
| Maximum pressure | 1200 bar |
|---|---|
| Maximum temperature | 90 °C |
| Pore size | 300 Å |
The recommended ratio is 1:20 (w/w).
It is desirable to incubate at 50 °C.
Yes, formic acid can be used as an alternative to TFA for protein digest analysis.