IntroductionQuality control (QC) is paramount in proteomics and biologics manufacturing and traditional methods can be limited by specificity and fragmentation behaviors; comprehensive sequence coverage is often lacking, especially for novel species or post-translational modifications (PTMs). Additionally, Multiple Attribute Monitoring (MAM) applied to different manufacturing stages and products requires robust tools that can handle different contaminants such as surfactants and stabilizers. To address these challenges, we present two innovative solutions: the Shredder, for in-depth, rapid proteomic sequencing with full sequence coverage, and a standardized, automatable MAM kit, based on the S-Trap Turbo 96-well plate, for streamlined biologics QC.MethodsThe Shredder applies a pseudo-enzymatic exceedingly low-specificity cleavage at all points along a peptide backbone to generate a series of highly overlapping peptides for complete sequence coverage. The MAM kit, based on a purpose-built version of the S-Trap Turbo, is tested under diverse conditions including surfactant-rich environments. The Shredder and MAM kit are applied to model proteins such as the NIST mAb RM8671 in the presence of contaminants like surfactants and polymers.Preliminary DataThe novel Shredder yielded unparalleled sequence coverage, producing an average of >10-100 peptides per amino acid position in a mere 30 minutes. This depth of analysis is akin to 'depth' in genomic sequencing and offers a granular view of the proteome, including elusive PTMs. We show the Shredder's efficiency compared to multiple enzymatic digestions on both model proteins and those of pharmaceutical interest, and subsequently propose possible applications in both academic and industry settings where verified PTM site localization and/or full sequence coverage may be essential. The S-Trap Turbo MAM kit was validated using the NISTmAb RM 8671 monoclonal antibody across challenging matrices, including viscous solutions and those with high surfactant content. It was robust in manual, semi-automated and fully automated workflows, and demonstrated high reproducibility including detection of PTMs and glycosylations, as well as residual host cell proteins (HCPs). This consistency was observed when comparing different automation platforms, with variations within the range of technical replicate injections. The standardized MAM kit worked successfully on samples with a myriad of contaminants and the Shredder provides unusually great sequence coverage. Together, they present a tool kit that address analytical needs at multiple stages of biopharmaceutical development. The technologies help ensure the detection of critical quality attributes to facilitating broad MAM adoption in therapeutic manufacturing from early research to final product release testing.Novel AspectFull-sequence peptide mapping and a universal MAM kit for standardized robust biotherapeutic QC.