NEW! We’re Launching IgMBRAZOR™!
IgMBRAZOR is a unique IgM-specific protease that digests human IgM at one specific site below the CH2 domain in the heavy chain, generating homogeneous F(ab’)2 and Fc fragments. We’re excited to launch this unique tool today that enables middle-level analysis of the complex and high molecular weight human IgM!
IgMBRAZOR facilitates IgM characterization during, for example, the development of vaccines, therapeutics, and diagnostics. The digestion is complete within 30 minutes, and due to the specificity of the enzyme, there is no risk of overdigestion if the incubation time is prolonged.
- Unique – enables middle-level characterization and quality control of IgM
- Fast – generates homogeneous F(ab’)2 and Fc fragments within 30 minutes
- Highly specific – one digestion site in the heavy chain of human IgM
Site-specific Digestion of IgM
IgMBRAZOR digests human IgM at one specific site below the CH2 domain, generating F(ab’)2 and Fc fragments, which allows for middle-level analysis of IgM. To demonstrate the site-specificity of IgMBRAZOR, human myeloma IgM was digested with IgMBRAZOR Lyophilized for 30 minutes at 37°C and compared to undigested IgM. To facilitate data interpretation the N-glycans were removed using PNGase F under denaturing and reducing conditions. The homogenous fragments observed in the mass spectra after IgMBRAZOR digestion and the masses of the detected fragments were consistent with digestion at one single site (..VPDQDT / AIRVFA..). The digestion using IgMBRAZOR is fast, easy, and carried out under physiological conditions, and the specificity of the enzyme prevents overdigestion of IgM and hydrolysis of other proteins, challenges otherwise commonly associated with other proteolytic enzymes. Middle-level analysis allows for the generation of high resolution mass data with minimal sample preparation and data analysis, which can be efficiently used for characterization, quality control, stability testing and production monitoring of therapeutic antibodies.
Site-specific digestion of IgM. IgM was digested with IgMBRAZOR Lyophilized for 30 minutes at 37°C, where after the Fc N-glycans were cleaved off using PNGase F Lyophilized under reducing and denaturing conditions to facilitate data interpretation. a) Schematic illustration of the sample preparation workflow. b) Deconvoluted mass spectra of the intact deglycosylated heavy chain and the VH-CH1-CH2 and CH3-CH4 fragments generated after IgMBRAZOR and PNGase F digestion. The inserts (top and bottom spectra) display the two peaks with and without the loss of the C-terminal tyrosine. The analysis was performed by reversed-phase LC-MS using a Waters™ BioAccord™ LC-MS system.
IgMBRAZOR Lyophilized – Process 1 mg human IgM.