Enhancing Negative-Ion Native Mass Spectrometry with Field-Assisted Capillary Vibrating Sharp-Edge Spray Ionization (cVSSI)

2021 ◽  
Author(s):  
Kushani Udeshika Attanayake
Keyword(s):  
Mass Spectrometry ◽  
Native Mass Spectrometry ◽  
Negative Ion ◽  
Sharp Edge

Rapid Online Buffer Exchange: A Method for Screening of Proteins, Protein Complexes, and Cell Lysates by Native Mass Spectrometry

2019 ◽  
Author(s):  
Zachary VanAernum ◽  
Florian Busch ◽  
Benjamin J. Jones ◽  
Mengxuan Jia ◽  
Zibo Chen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Speed ◽  
Structural Information ◽  
Protein Complexes ◽  
High Sensitivity ◽  
Native Mass Spectrometry ◽  
Structural Features ◽  
Consumer Products ◽  
Cell Lysates ◽  
Protein Expression And Purification

It is important to assess the identity and purity of proteins and protein complexes during and after protein purification to ensure that samples are of sufficient quality for further biochemical and structural characterization, as well as for use in consumer products, chemical processes, and therapeutics. Native mass spectrometry (nMS) has become an important tool in protein analysis due to its ability to retain non-covalent interactions during measurements, making it possible to obtain protein structural information with high sensitivity and at high speed. Interferences from the presence of non-volatiles are typically alleviated by offline buffer exchange, which is timeconsuming and difficult to automate. We provide a protocol for rapid online buffer exchange (OBE) nMS to directly screen structural features of pre-purified proteins, protein complexes, or clarified cell lysates. Information obtained by OBE nMS can be used for fast (<5 min) quality control and can further guide protein expression and purification optimization.

Confounding contaminants in mass spectrometric reaction monitoring

2018 ◽  
Author(s):  
Gilian T. Thomas ◽  
Landon MacGillivray ◽  
Natalie L. Dean ◽  
Rhonda L. Stoddard ◽  
Lars Yunker ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Mass Spectrometric ◽  
Negative Ion ◽  
Reaction Monitoring ◽  
Schlenk Flask ◽  
Dynamic Analyses ◽  
Rubber Septa ◽  
Negative Ion Mode ◽  
Mode A

<p>Reactions carried out in the presence of rubber septa run the risk of additives being leached out by the solvent. Normally, such species are present at low enough levels that they do not interfere with the reaction significantly. However, when studying reactions using sensitive methods such as mass spectrometry, the appearance of even trace amounts of material can confuse dynamic analyses of reactions. A wide variety of additives are present in rubber along with the polymer: antioxidants, dyes, detergent, and vulcanization agents, and these are all especially problematic in negative ion mode. A redesigned Schlenk flask for pressurized sample infusion (PSI) is presented as a means of practically eliminating the presence of contaminants during reaction analyses.</p>

Native Mass Spectrometry-Based Screening for Optimal Sample Preparation in Single Particle Cryo-EM

2020 ◽  
Author(s):  
Paul Dominic B. Olinares ◽  
Jin Young Kang ◽  
Eliza Llewellyn ◽  
Courtney Chiu ◽  
James Chen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Sample Preparation ◽  
Single Particle ◽  
Native Mass Spectrometry ◽  
Optimal Sample
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