Oct 15
We present here a software suite (ORIGAMI) that facilitates the rapid acquisition and analysis of ion mobility data following collisional activation. ORIGAMI was developed for use on Waters Synapt instruments where data acquisition is achieved by interfacing WREnS (Waters Research Enabled Software) and MassLynx. Two components are presented, the first is ORIGAMIMS which enables the activation of ions via a …
Mar 02
The aggregation of protein-based therapeutics such as monoclonal antibodies (mAbs) can affect the efficacy of the treatment and can even induce effects that are adverse to the patient. Protein engineering is used to shift the mAb away from an aggregation-prone state by increasing the thermodynamic stability of the native fold, which might in turn alter …
Oct 09
Proteins and protein complexes undergo structural changes depending on the surrounding environment, in turn affecting their biological function. Here, VT-MS is applied to study four multimeric protein complexes and allow for decoupling of their melting temperature (Tm) from the protein complex dissociation temperature (TGPD). VT-IM-MS is used to investigate structural changes of these proteins at …
Oct 09
Monoclonal antibodies (mAb) are a rapidly growing group of biopharmaceuticals. Due to their intrinsic flexibility, intact mAbs provide a challenge with regards to higher order structure characterization. Intact mAbs and their fragments are explored here using the DT-IM-MS and were found to be far more dynamic in the gas-phase than proteins of comparable size. This …
Oct 08
Thermally induced conformational transitions of three proteins of increasing intrinsic disorder—cytochrome c, the tumor suppressor protein p53 DNA binding domain (p53 DBD), and the N-terminus of the oncoprotein murine double minute 2 (NT-MDM2)—have been studied by native mass spectrometry and variable-temperature drift time ion mobility mass spectrometry (VT-DT-IM-MS). Ion mobility measurements were carried out at temperatures …
Oct 08
Developing drug-like molecules to inhibit the interactions formed by disordered proteins is desirable due to the high correlation of disorder with protein implicated in disease, but is challenging due in part to the lack of atomistically resolved and resolvable structures from conformationally dynamic systems. Ion mobility mass spectrometry (IM-MS) is well-positioned to assess protein ligand …
Oct 08
An ion mobility mass spectrometer has been modified to allow optical interrogation of ions with different mass-to-charge (m/z) ratios and/or mobilities (K). An ion gating and trapping procedure has been developed which allows us to store ions for several seconds enabling UV photodissociation (UVPD). The instrument has been modified to enable overlap of the laser …
