Intact mAb Mass Check Standard
The Intact mAb Mass Check Standard is a meticulously characterized monoclonal antibody (mAb) reference standard designed to facilitate the qualitative analysis of intact proteins via liquid chromatography-mass spectrometry (LC-MS). This standard is instrumental in protein identification, mAb characterization, and the detection of post-translational modifications, ensuring that mass spectrometry systems are optimally tuned and calibrated for accurate, high-resolution measurements.
Derived from an intact mouse immunoglobulin G1 (IgG1) protein purified through Protein-A affinity chromatography, the Intact mAb Mass Check Standard possesses a well-defined molecular weight of 145,329.7 Daltons. Its molecular composition is detailed as C6472 H9940 N1698 O2008 S52, featuring 17 disulfide bonds that maintain structural stability. Notably, each heavy chain's N-terminus undergoes pyroglutamic acid modification, and there is a single N-linked glycosylation site per heavy chain, resulting in four primary glycoforms: G0, G0F, G1F, and G2F. This precise molecular characterization makes the standard an invaluable tool for assessing instrument performance and method development in intact mass analysis.
In practical applications, the Intact mAb Mass Check Standard serves as a reliable reference for routine mass spectrometry optimization, system setup, and performance verification. Providing a consistent and reproducible benchmark enables laboratories to fine-tune their LC-MS systems, ensuring accurate mass measurements and robust detection of protein modifications.
This is particularly crucial in biopharmaceutical development, where understanding the molecular profile of therapeutic antibodies is essential for efficacy and safety assessments.
For users of the GlycoWorks RapiFluor-MS N-Glycan sample preparation workflow, the Intact mAb Mass Check Standard functions as an unlabeled control or reference standard. Its four major glycosylated peptides mirror those commonly encountered in therapeutic mAbs, providing a relevant benchmark for method validation and comparative studies. This alignment ensures that glycan analysis workflows yield accurate and reliable data, facilitating the characterization of glycosylation patterns critical to mAb function and stability.
The standard is supplied as 1 mg of lyophilized powder, stabilized with lyophilization-grade sucrose, and packaged in Waters TruView Vials. These vials are specifically designed to minimize analyte adsorption, preserving sample integrity, especially for peptide and protein analyses. Upon reconstitution, the standard provides a stable and ready-to-use solution for immediate integration into LC-MS workflows.
The Intact mAb Mass Check Standard is an essential resource for laboratories engaged in protein analysis and monoclonal antibody characterization. Its comprehensive molecular definition, coupled with its role in system calibration and method validation, supports the generation of precise and reproducible data in intact mass spectrometry applications. By incorporating this standard into their analytical protocols, scientists can enhance the reliability and accuracy of their protein analyses, thereby advancing research and development in the biopharmaceutical field.
FAQs
What are the key molecular properties of the Intact mAb Mass Check Standard?
The Intact mAb Mass Check Standard is a fully characterized mouse IgG1 monoclonal antibody with a precise molecular weight of 145,329.7 Da. Its molecular formula is C6472 H9940 N1698 O2008 S52, reflecting its complex structure. It contains 17 disulfide bonds, which contribute to structural stability. The N-terminus of each heavy chain features a pyroglutamic acid modification, a common post-translational modification in mAbs. Additionally, each heavy chain has one N-linked glycosylation site, resulting in four major glycoforms: G0, G0F, G1F, and G2F. These properties make it a reliable reference for LC-MS system calibration and intact protein mass analysis.
How is the Intact mAb Mass Check Standard characterized?
The Intact mAb Mass Check Standard is fully characterized by rigorous analytical testing, which ensures it serves as a reliable LC-MS reference standard. Its precise molecular weight (145,329.7 Da) and defined glycoform composition provide a consistent benchmark for mass spectrometry analysis. It is purified using Protein-A chromatography, ensuring high purity and reproducibility.
The standard's 17 disulfide bonds, pyroglutamic acid modification at the N-terminus, and single N-linked glycosylation site per heavy chain further enhance its characterization. Because of these well-defined structural attributes, it is ideal for optimizing mass spectrometer performance, system calibration, and protein modification analysis in biopharmaceutical research.
Intact mAb Mass Check Standard
