ABSTRACT
Protein-based biopharmaceutical drugs, such as monoclonal antibodies, account for the majority of the best-selling drugs globally in recent years. For bioprocesses, key performance indicators are the concentration and aggregate level for the product being produced. In water NMR (wNMR), the use of the water transverse relaxation rate [R2(1H2O)] has been previously used to determine protein concentration and aggregate level; however, it cannot be used to separate between them without using an additional technique. This work shows that it is possible to "decouple" these two key characteristics by recording the water diffusion coefficient [D(1H2O)] in conjunction with R2(1H2O), even in the event of overlap in either D(1H2O) or R2(1H2O). This method is demonstrated on three different systems, following appropriate D(1H2O) or R2(1H2O) calibration data acquisition for a protein of interest. Our method highlights the potential use of benchtop NMR as an at-line process analytical technique.
Subject(s)
Water , Water/chemistry , Diffusion , Nuclear Magnetic Resonance, Biomolecular/methods , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/analysis , Proteins/analysis , Proteins/chemistry , Protein Aggregates , Magnetic Resonance Spectroscopy/methodsABSTRACT
We report the first total synthesis of samroiyotmycinâ A (1), a C2 -symmetric 20-membered anti-malarial macrodiolide isolated from Streptomyces sp. The convergent synthetic strategy orchestrates bisalkyne fragment-assembly using an unprecedented Schöllkopf-type condensation on a substituted ß-lactone and an ambitious late-stage one-pot alkyne cross metathesis-ring-closing metathesis (ACM-RCAM) reaction. The demanding alkyne metathesis sequence is achieved using the latest generation of molybdenum alkylidynes endowed with a tripodal silanolate ligand framework. Subsequent conversion to the required E-alkenes uses contemporary hydrometallation chemistry catalysed by tetrameric cluster [{Cp*RuCl}4 ].