Maxis II

Middle-Up and Middle-Down Antibody Subunit Analysis

Subunit analysis delivers fast confirmation of expected domain molecular weights, sample heterogeneity and sequence validation.

Subunit analysis provides a rapid and simple screening method for biotherapeutic antibodies that delivers critical information for process development while requiring minimal sample prep and data analysis. This enables the determination of multiple quality attributes at early stages when detailed LCMS peptide maps would be cost prohibitive.

Adulimumab heavy chain analysed on the maXis
Adulimumab heavy chain analysed on the maXis II. The isotopically resolved deconvoluted data (blue) shows strong alignment with the expected isotopic pattern (black) which allows calculation of the monoisotopic mass at better than 1 ppm mass accuracy

With up to 80,000 resolution, sub ppm mass accuracy, True Isotopic Patterns & industry leading sensitivity at the subunit level, Bruker UHR-QTOFs provide confident characterization first time, every time. Isotopic resolution of antibody subunits allows clear interpretation of protein modifications and routinely delivers monoisotopic masses at better than 2ppm mass accuracy. Bruker ESI-QTOFs offer a middle-up solution enabling UHPLC speed analysis with no sensitivity or resolution compromises, uniquely allowing simultaneous characterization of product & impurities in a single analysis.

ESI-QTOF middle-up analysis is easily complemented by adding sequence information via middle-down experiments, which also benefits from the same minimal sample preparation. The maXis II ETD can provide confirmation of N- and C-terminal sequences, whilst LC-MALDI can be used for validation of the full subunit sequences.  These approaches can be supplemented with intact mass and middle-up measurements to provide conclusive information for sequence variant detection and biosimilar development.