Capillary Electrophoresis-Mass Spectrometry: Methods and Protocols (Methods in Molecular Biology, 2531)

264 Pages·2022·10.781 MB·English

by  Christian Neusüß (editor), Kevin Jooß (editor)

Methods in
Molecular Biology 2531
Christian Neusüß · Kevin Jooß Editors
Capillary
Electrophoresis-
Mass Spectrometry
Methods and Protocols
M M B
ETHODS IN OLECULAR IO LO GY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire, UK
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Capillary Electrophoresis-Mass
Spectrometry
Methods and Protocols
Edited by
Christian Neusüß and Kevin Jooß
Aalen University, Aalen, Baden-Württemberg, Germany
Editors
ChristianNeusu¨ß KevinJooß
AalenUniversity AalenUniversity
Aalen,Baden-Wu¨rttemberg,Germany Aalen,Baden-Wu¨rttemberg,Germany
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-0716-2492-0 ISBN978-1-0716-2493-7 (eBook)
https://doi.org/10.1007/978-1-0716-2493-7
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Preface
Capillary electrophoresis-mass spectrometry (CE-MS) is still an underutilized technique
compared to the widespread liquid chromatography-mass spectrometry (LC-MS). Never-
theless,theselectivitymakesCE-MSanattractivetoolinmanyfieldsofapplicationwhichis
oftentimes considered complementary to traditional LC separation. In general, electromi-
grativetechniquesseparateionsbasedontheircharge-to-sizeratioand/orisoelectricpoint.
Consequently, high separation efficiency can be achieved, ideally resulting in exceptionally
sharp peaks under diffusion-limited conditions. In this way, it is possible to resolve closely
related species such as isomeric molecules (including enantiomers) or proteoforms varying
inonlyoneposttranslationalmodification.
CErequirestheapplicationofhighelectricfieldstoperformreasonablyfastandefficient
separations, resulting in high electric currents. Consequently, efficient heat dissipation is
requiredtoavoidpeakbroadeningandbubbleformationduetoJouleheating.Thisleadsto
smallgeometrieswithrelativelyhighsurfaceresultingintotalcapillaryvolumestypicallyin
thesingledigitmicroliterrange.Thus,CEisamicrofluidicseparationtechniqueconsidered
environmentally friendly due to its low consumption of resources (e.g., solvents). For the
samereason,CE-MSisparticularlysuitablefor theanalysisofvaluablesamples,e.g.,ifonly
minor amounts are available. However, on the other hand, analytes need to be sufficiently
concentrated for sensitive detection. In addition, the handling of small volumes requires
specialcareandcanbetechnicallychallenging.
Mass Spectrometry (MS) acts as a universal and selective technique for molecule
characterization and can be applied in both targeted and nontargeted fashion. In the last
decades, enormous progress has been made in MS technology leading to high sensitivity,
high speed, increased resolving power, the ability for selective fragmentation of small and
largemolecules,andlastbutnotleastimprovedoverallusability.Still,separationtechniques
arecrucialduetopossibleinterferenceduringtheionizationprocessandmassspectrometric
resolution being limited, which is especially important in the context of complex samples.
ElectrosprayIonization(ESI)isthetechniqueofchoiceforon-linecouplingofliquidphase
separations, which is particularly true for electromigrative techniques. However, the need
for closing the electric circuits for both the CE and the ESI process at the end of the
separation capillary requires unique interface designs. Moreover, the selectivity of electro-
migrative techniques is substantially influenced by the type and concentration of the com-
ponents ofthebackground electrolyte (BGE).ManyCEmethodsarenot compatiblewith
ESI-MS in a direct fashion due to nonvolatile constituents, being required for best separa-
tionperformance.ThesespecialneedsforCE-MSareaddressedinthisbook.
In the first part the handling of modern interfaces is described in detail, including
approaches based on (nano)sheath liquid (Chapters 1 and 2), porous tip (Chapter 3), and
liquid junction CE-MS interfaces (Chapter 4). The development of novel and increasingly
sophisticatedcapillarycoatingsisconsideredanimportantpartofCE-MSmethoddevelop-
ment. In this regard, a protocol on an efficient coating for protein analysis is included (see
Chapter5).CEtechnologyallowstheintegrationofshortsectionscontainingsolidmaterial
(e.g., functionalized beads) for specific and efficient manipulation of analyte molecules.
Therefore, one chapter was dedicated to utilizing an immobilized enzyme reactor imple-
mented in the CE capillary for tryptic digestion of intact proteins and subsequent peptide
v
vi Preface
level analysis (see Chapter 6). This chapter is followed by a CE-MS-dedicated protocol for
the sample preparation and analysis of tryptic peptides (see Chapter 7). Intact protein
characterization is another important field of CE-MS applications. In this context, top-
down proteomics (TDP) gets increasing attention due to its capability to circumvent the
inference problem in proteomics. A general TDP protocol for CE-MS can be found in
Chapter 8. As a representative of the field of biotherapeutics the analysis of monoclonal
antibodies,rangingfromreducedtointactlevel,issubsequentlydescribed(seeChapter9).
ThecomplexityofglycoproteinscanbequitechallengingwithCE-MSbeinghighlysuited
toseparateglycoforms.ACE-MSmethoddedicatedtobottom-upanalysisofglycoproteins
is presented in Chapter 10. Another protocol is dedicated to the characterization of
glycosaminoglycans, more specifically, the chondroitin sulfate and dermatan sulfate, which
i.a. play an essential role at the extracellular matrix (see Chapter 11). Two protocols on
metaboliteanalysisforcations(seeChapter12)andanions(seeChapter13)showthestellar
powerofCE-MSinthisimportantfield.CEinelectrokineticchromatographymodeiswell
suited for chiral analysis and described in detail for four different fields of application
including pharmaceutical, biomedical, food, and agrochemical analysis (see Chapter 14).
Traditionally, CE has been ruled out of many food safety applications, due to the lack of
mandatory sensitivity. The inclusion of chromatography-based approaches for sample
enrichmenthasbeenthemostsuccessful.Aprotocolutilizingin-linesolidphaseextraction
for the detection of quinolone antibiotic residues in chicken meat is presented in
Chapter15.AlesscommonsetupisthehyphenationofCEwithinductivelycoupledplasma
massspectrometry(ICP-MS).Ithasbeendemonstrated thatthiscombinationisespecially
suitableforthecharacterizationofmetalnanoparticles,beingitintraditionalICP-MSmode
orevensingleparticle(sp)ICP-MS(seeChapter16).
Overall,thisbookisdedicatedtohelpnormalizeCE-MSpracticesandprovideguidance
coveringavarietyofdifferentfieldsofapplicationrangingfromsmallmoleculesuptolarge
intact proteins. We hope to enhance the CE-MS community and supply a resource for
production of reproducible and high-quality data. We thank all authors for their valuable
contribution and their patience in the editorial process. We would also like to thank the
Methods in Molecular Biology series editor, John Walker, for his guidance and impelling
motivationinpreparingthisvolume.
Aalen,Baden-Wu¨rttemberg,Germany ChristianNeusu¨ß
KevinJooß
Contents
Preface ..................................................................... v
Contributors................................................................. ix
1 EvaluationofPrototypeCE-MSInterfaces................................. 1
SabrinaFerre´,JulienBoccard,SergeRudaz,andVı´ctorGonza´lez-Ruiz
2 CapillaryElectrophoresis–MassSpectrometry(CE-MS)by
Sheath–FlowNanosprayInterfaceandItsUsein
BiopharmaceuticalApplications........................................... 15
MeiHan,RichardSmith,andDanA.Rock
3 ProtocolforEtchingBare-FusedSilicaCapillariesforSheathless
CapillaryElectrophoresis–MassSpectrometryCoupling ..................... 49
ClarisseGosset-Erard,Je´re´mieGiorgetti,MichaelBiacchi,
Fre´de´ricAubriet,EmmanuelleLeize-Wagner,
PatrickChaimbault,andYannis-NicolasFranc¸ois
4 MicrofabricatedLiquidJunctionCapillaryElectrophoresis–Mass
SpectrometryInterface .................................................. 61
JanaKrenkovaandFrantisekForet
5 SuccessiveMultipleIonic-PolymerLayerCoatingsforIntact
ProteinAnalysisbyCapillaryZoneElectrophoresis–MassSpectrometry:
ApplicationtoHemoglobinAnalysis ...................................... 69
LiesaSalzer,AlexanderStolz,LauraDhellemmes,AlisaH¨ochsmann,
LaurentLeclercq,Herve´Cottet,andChristianNeusu¨ß
6 On-lineImmobilizedEnzymeMicroreactorCapillaryZone
Electrophoresis–MassSpectrometryforPeptideMapping.................... 77
RogerPero-Gascon,LauraPont,EstelaGime´nez,
VictoriaSanz-Nebot,andFernandoBenavente
7 Bottom-UpAnalysisofProteinsbyPeptideMassFingerprinting
withtCITP-CZE-ESI-TOFMSAfterTrypticDigest........................ 93
MariusSeglandHannoStutz
8 Top-DownProteomicsbyCapillaryZoneElectrophoresis-Tandem
MassSpectrometryforLarge-ScaleCharacterizationofProteoforms
inComplexSamples..................................................... 107
ElijahN.McCool,RacheleA.Lubeckyj,DaoyangChen,
andLiangliangSun
9 AssessmentofMacro-andMicroheterogeneityofMonoclonal
AntibodiesUsingCapillaryZoneElectrophoresisHyphenated
withMassSpectrometry ................................................. 125
ChristophGst¨ottner,RobHaselberg,ManfredWuhrer,
GovertW.Somsen,andElenaDomı´nguez-Vega
10 High-SensitivityGlycoproteomicAnalysisofBiological
SamplesbyCZE-ESI-MS................................................ 143
W.WangandG.S.M.Lageveen-Kammeijer
vii
viii Contents
11 CapillaryZoneElectrophoresis–ElectrosprayIonizationTandem
MassSpectrometryforTotalAnalysisofChondroitin/Dermatan
SulfateOligosaccharides ................................................. 163
AlinaD.Zamfir
12 In-SourceFragmentationfor theIdentificationofCompounds
byCE-ESI-TOFinHumanPlasma.L-ProlineasCaseStudy ................. 185
MaricruzMamani-Huanca,AnaGradillas,
A´ngelesL(cid:1)opez-Gonza´lvez,andCoralBarbas
13 AssessingtheEnergyStatusofLowNumbersofMammalian
CellsbyCapillaryElectrophoresis–MassSpectrometry....................... 203
WeiZhangandRawiRamautar
14 ApplicationsofCapillaryElectrophoresis-MassSpectrometry
toChiralAnalysis ....................................................... 211
ElenaSa´nchez-L(cid:1)opezandMarı´aLuisaMarina
15 ImprovedSensitivitytoDetermineAntibioticResiduesin
ChickenMeatbyIn-LineSolid-PhaseExtractionCoupled
toCapillaryElectrophoresis–TandemMassSpectrometry.................... 227
FranciscoJ.LaraandA.M.Garcı´a-Campan˜a
16 CECoupledtoICP-MSandSingleParticleICP-MSfor
NanoparticleAnalysis.................................................... 243
DaryaMozhayevaandCarstenEngelhard
Index ...................................................................... 259
Contributors
FRE´DE´RIC AUBRIET • LaboratoiredeChimieetPhysique-ApprocheMulti-e´chellesdesMilieux
Complexes(LCP-A2MC),Universite´deLorraine,Metz,France
CORALBARBAS • CentrodeMetabol(cid:1)omicayBioana´lisis(CEMBIO),FacultaddeFarmacia,
UniversidadSanPablo-CEU,CEUUniversities,Urbanizaci(cid:1)onMonteprı´ncipe,Madrid,
Spain
FERNANDOBENAVENTE • DepartmentofChemicalEngineeringandAnalyticalChemistry,
InstituteforResearchonNutritionandFoodSafety(INSAlUB),UniversityofBarcelona,
Barcelona,Spain
MICHAELBIACCHI • LaboratoiredeSpectrome´triedeMassedesInteractionsetdesSyste`mes
(LSMIS),UMR7140(Unistra-CNRS),Universite´deStrasbourg,Strasbourg,France
JULIENBOCCARD • InstituteofPharmaceuticalSciencesofWesternSwitzerland(ISPSO),
UniversityofGeneva,Geneva,Switzerland;SchoolofPharmaceuticalSciences,University
ofGeneva,Geneva,Switzerland;SwissCentreforAppliedHumanToxicology,Basel,
Switzerland
PATRICKCHAIMBAULT • LaboratoiredeChimieetPhysique-ApprocheMulti-e´chellesdes
MilieuxComplexes(LCP-A2MC),Universite´deLorraine,Metz,France
DAOYANGCHEN • DepartmentofChemistry,MichiganStateUniversity,EastLansing,MI,
USA
HERVE´ COTTET • InstitutdesBiomole´culesMaxMousseron,UMRCNRS,Universite´de
Montpellier,Montpellier,France
LAURADHELLEMMES • InstitutdesBiomole´culesMaxMousseron,UMRCNRS,Universite´de
Montpellier,Montpellier,France
ELENADOMI´NGUEZ-VEGA • Center forProteomicsandMetabolomics,LeidenUniversity
MedicalCenter,Leiden,TheNetherlands
CARSTENENGELHARD • DepartmentofChemistryandBiology,UniversityofSiegen,Siegen,
Germany;ResearchCenterofMicro-andNanochemistryand(Bio)Technology,University
ofSiegen,Siegen,Germany
SABRINAFERRE´ • InstituteofPharmaceuticalSciencesofWesternSwitzerland(ISPSO),
UniversityofGeneva,Geneva,Switzerland;SchoolofPharmaceuticalSciences,University
ofGeneva,Geneva,Switzerland
FRANTISEKFORET • InstituteofAnalyticalChemistryoftheCzechAcademyofSciences,Brno,
CzechRepublic
YANNIS-NICOLASFRANC¸OIS • LaboratoiredeSpectrome´triedeMassedesInteractionsetdes
Syste`mes(LSMIS),UMR7140(Unistra-CNRS),Universite´deStrasbourg,Strasbourg,
France
A.M.GARCI´A-CAMPAN˜A • DepartmentofAnalyticalChemistry,FacultyofSciences,
UniversityofGranada,Granada,Spain
ESTELA GIME´NEZ • DepartmentofChemicalEngineeringandAnalyticalChemistry,
InstituteforResearchonNutritionandFoodSafety(INSAlUB),UniversityofBarcelona,
Barcelona,Spain
JE´RE´MIEGIORGETTI • LaboratoiredeSpectrome´triedeMassedesInteractionsetdesSyste`mes
(LSMIS),UMR7140(Unistra-CNRS),Universite´deStrasbourg,Strasbourg,France
ix