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Influence of drople...
Influence of droplet size, capillary-cone distance and selected instrumental parameters for the analysis of noncovalent protein-ligand complexes by nano-electrospray ionization mass spectrometry
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- Benkestock, Kurt (author)
- KTH,Kemi
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- Sundqvist, Gustav (author)
- KTH,Kemi
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- Edlund, Per Olof (author)
- Biovitrum AB, Dept Analyt Sci
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- Roeraade, Johan (author)
- KTH,Kemi
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(creator_code:org_t)
- 2004-09-24
- 2004
- English.
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In: Journal of Mass Spectrometry. - : Wiley. - 1076-5174 .- 1096-9888. ; 39:9, s. 1059-1067
- Related links:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
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- It has been suggested in the literature that nano-electrospray ionization (nano-ESI) mass spectrometry better reflects the equilibrium between complex and free protein in solution than pneumatically assisted electrospray ionization (ESI) in noncovalent interaction studies. However, no systematic studies of the effects of ionization conditions have been performed to support this statement. In the present work, different instrumental and sample-derived parameters affecting the stability of noncovalent complexes during analysis by nano-ESI were investigated. In general, increased values of parameters such as drying gas flow-rate, ion-source temperature, capillary tip voltage and buffer concentration lead to a dissociation of ribonuclease A (RNAse)-cytidine 2'-monophosphate (CMP) and cytidine 5'-triphosphate (CTP) complexes. The size of the electrosprayed droplets was shown to be an important issue. Increasing the capillary to cone distance yielded an increased complex to free protein ratio when a hydrophilic ligand was present and the reverse effect was obtained with a hydrophobic ligand. Important in this regard is the degree of sampling of ions originating from late-generation residue droplets, that is, ions present in the droplet bulk. Sampling of these ions increases with longer capillary-cone distance (flight time). Furthermore, when the sample flow-rate was increased by increasing the capillary internal tip i.d. from 4 to 30 mum, a decreased complex to free protein ratio for the RNAse-CTP system was observed. This behavior was consistent with the change in surface to volume ratio for flow-rates between 2 and 100 nl min(-1). Finally, polarity switching between positive and negative ion modes gave a higher complex to free protein ratio when the ligand and the protein had the same polarity.
Subject headings
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
Keyword
- nano-electrospray ionization; noncovalent complexes; droplet size; capillary-cone distance; H-FABP; RNAse; BINDING INTERACTIONS; QUANTITATIVE-DETERMINATION; COMPETITIVE-BINDING; RESOLUTION; RECEPTOR; CHARGE; DNA
- Analytical chemistry
- Analytisk kemi
Publication and Content Type
- ref (subject category)
- art (subject category)
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