Intact Protein Separations: Humanized mAb Mass Check Standard

The analysis of intact proteins by LC-MS is a preferred approach for the characterization of biopharmaceuticals, in particular monoclonal antibodies (mAbs). In such an analysis, it is possible to acquire intact mass information, quickly confirm sequence and post-translation modifications, as well as interrogate product related impurities. To support this application area, Waters has commercialized the Humanized mAb Mass Check Standard, which can be used in the benchmarking of LC-MS techniques, proficiency testing among different instruments and laboratories, and system suitability when doing intact protein analysis. This standard is a stabilized and lyophilized format of NIST Reference Material 8671 (NIST mAb), a humanized IgG1κ expressed from a murine cell line.*

*Disclaimer: Reference values for RM 8671 are not certified to be in correspondence with any Report of Investigation offered by the National Institute of Standards and Technology (NIST). If it is desirable to have such correspondence, it is advised that RM 8671 be directly acquired from NIST (nist.gov).

Reversed-Phase Protein Separations: The MassPREP Protein Standard Mix

This Intact Protein Validation Mix is designed for verification of HPLC-UPLC instruments. This standard is also used to evaluate the performance of columns used for protein analysis, specifically the ACQUITY UPLC and XBridge Protein BEH C₄ 300Å Columns. The MassPrep Protein Standard contains proteins that have a wide range of isoelectric points, molecular weights, and hydrophobicities.  

Size Exclusion Separations: SEC Protein Standard Mixes

Whether using UPLC or HPLC, the SEC Protein Standard Mixes are unique, specially formulated protein mixtures specific to the BEH 200Å, BEH 125Å or BEH 450Å columns.  It is recommended that they be used on a regular basis for evaluation of these types of columns for optimal system performance.

Ion Exchange Separations: IEX Test Standards

These standards are specially designed for either cation or anion exchange chromatography and each provide a unique set of 3 proteins found to provide good chromatographic separation based on the charge state being utilized.

System Check: Complex Protein Standards

Analysis of intact proteins by MS is a powerful approach for protein identification and assignment of protein modifications. The Intact mAb Mass Check Standard is a LC-MS standard used as a qualitative tool for intact mass analysis. This standard is designed to be used for routine MS optimization, system setup and performance testing. It is an intact mouse IgG1 protein purified by Protein-A with a known molecular weight, ideal for higher molecular weight intact mass measurements.  It is provided in a Waters TruView Vial (ideal for peptides) as 1 mg of lyophilized solid (lyophilization grade sucrose is used for stabilization). Recommended solubilization is in 100-200 µL of DI water.  Store at -20 °C immediately upon arrival.