Best practices and benchmarks for intact protein analysis
In this paper we provide a decision tree that guides researchers to robust protocols for mass analysis of intact proteins (antibodies, membrane proteins and others) from mixtures of varying complexity. We also present cross-platform analytical benchmarks using a protein standard sample, to allow users to gauge their proficiency.
Top-down MS of native proteoforms and their complexes
A study to develop and test protocols for native MS combined with top-down fragmentation of proteins and protein complexes across 11 instruments in nine laboratories. Here we report the summary of the outcomes to provide robust benchmarks and a valuable entry point for the scientific community.
Top-Down MS and MS Methods for Bruker Instruments
This documents shows standard methods for protein mixture preparation, separation by LC as well as parameters for successful MS and MS measurements. These parameters are not set for ultimate performance but rather offer a dependable starting point to help you become successful in your research.
Top-Down MS and MS Methods for Thermo Instruments
This documents shows standard methods for protein mixture preparation, separation by LC as well as parameters for successful MS and MS measurements. These parameters are not set for ultimate performance but rather offer a dependable starting point to help you become successful in your research.
Fragmentation of the hinge region of IgG, producing a F(ab’)2 fragment and two Fc fragments
FabRICATOR® is a digestive enzyme that cleaves IgG only at one specific site below the hinge region resulting in F(ab’)2 and Fc-fragments. Since FabRICATOR® only cleaves at one specific site below the hinge region, there is no risk of over digestion if the incubation time is increased for several hours.
Fragmentation of human IgG1 between K223 and T224 producing a homogenous pool of Fab and Fc fragments
GingisKHAN™ digests human IgG1 at one single site in the upper hinge (…KSCDK / THTCPPCP…). A second cleavage site on the Fc may appear if the N-glycans are removed. GingisKHAN™ is a cysteine protease and requires reducing environment to be active. Intact Fab and Fc fragments are obtained with GingisKHAN™ digestion of human IgG1 since mild reducing conditions (i.e. 2 mM cysteine) is sufficient for enzyme activity.
Human Plasma Sample Preparation Workflow Using the Seer Proteograph
This document demonstrates the method of using engineered nanoparticles for sampling plasma proteins across the wide dynamic range of the proteome enabling biomarker discovery.