ELISA: Methods and Protocols

245 Pages·2023·11.302 MB·English

by  Robert Matson

Methods in
Molecular Biology 2612
Robert S. Matson
Editor
ELISA
Methods and Protocols
M M B
ETHODS IN OLECULAR IO LO GY
SeriesEditor
JohnM.Walker
School of Lifeand MedicalSciences
University ofHertfordshire
Hatfield, Hertfordshire, UK
Forfurther volumes:
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ELISA
Methods and Protocols
Edited by
Robert S. Matson
QuantiScientifics LLC, Orange, CA, USA
Editor
RobertS.Matson
QuantiScientificsLLC
Orange,CA,USA
ISSN1064-3745 ISSN1940-6029 (electronic)
MethodsinMolecularBiology
ISBN978-1-0716-2902-4 ISBN978-1-0716-2903-1 (eBook)
https://doi.org/10.1007/978-1-0716-2903-1
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Dedication
for
LielleandAyla
Preface
This edition of ELISA: Methods and Protocols was written during the challenging times
associatedwiththeonslaughtoftheCovid-19worldwidepandemic.Itistrulyremarkableto
havewitnessedtherapidemergenceoftherecombinant(mRNA/DNA)vaccinestrategyto
combat this formidable enemy as well as others we have yet to know. We must also
acknowledge the tremendous effort of the diagnostic industry to provide reliable testing
on such a scale and with very short notice. In the backdrop, there was an army of skilled
scientists supporting these efforts.It is very likelythat manyused theELISA in support of
theirimportantwork.
Whiletheenzyme-linkedimmunosorbentassay(ELISA)isoneofthemostwidelyused
techniques, its nuances are not always fully understood. Case in point, the revelation that
excessivebiotinconsumptionmayledtoassayinterferencewhenusingthewell-established
streptavidin-biotinreportersystem.Youwillfindthattheauthorsoffergreatinsightintothe
theory and practice of the ELISA with useful notes to help you through the step-by-step
protocolswhilesteeringyouclearofcommonpitfalls.
In this compendium, with the introductory chapter we start with a review of the basic
formatsusedintheELISA,theirapplication,strengths,andweaknesses.Thisisfollowedby
examining the essential components and features comprising the technique-enzyme, anti-
body, and sorbent. Approaches to the immobilization of antibodies are provided for those
who may find a need to prepare their own immunosorbent. Since the antibody specificity
andsensitivityaremostcriticaltothesuccessoftheassay,achapterisdedicatedtoscreening
andselectionofantibodies.Next,wediscussmeanstoeffectivelyblockthesorbentsurface
toreducenon-specificbinding.Inanotherchapter,theadvancesinsurfacemodificationof
the plastics commonly used as the sorbent are described. A chapter on interference com-
pletesthefirstpartofthebook.
In the second half of the book, we highlight applications of the ELISA. These include
the determination of cytokines in tissues and cerebral spinal fluid by sandwich ELISA, a
microarray-based method for carbohydrate characterization, and a chapter on mycotoxin
extractionwithdeterminationbycompetitiveimmunoassay.AutomationofELISAforhigh-
throughputdiagnosticapplications,multiplexallergentesting,andadaptationofELISAfor
lateral flow are described in other chapters. There is exciting work on a homogeneous
immunoassay and a final chapter examining the role of ELISA in biosensors. It is the goal
ofthisbooktoprovidethereaderwithabasicunderstandingofthetechniquewithmethods
andprotocolsthatenablesuccessatperformingyourenzyme-linkedimmunosorbentassay.
Orange,CA,USA RobertS.Matson
vii
Contents
Dedication.................................................................. v
Preface ..................................................................... vii
Contributors................................................................. xi
1 Enzyme-LinkedImmunosorbentAssay:TypesandApplications...... ........ 1
HovhannesHayrapetyan,ThaoTran,EglisTellez-Corrales,
andCharithaMadiraju
2 ELISAEssentials:Surfaces,Antibodies,Enzymes,andSubstrates..... ........ 19
RobertS.Matson
3 AntibodyImmobilization........... ....... ....... ........ ....... ........ 33
RobertS.Matson
4 ScreeningforAntibodySpecificityandSensitivitywithELISA........ ........ 45
MichaelJ.HalatandEmilyGravens
5 EffectiveBlockingandStabilizingMethodsUsingSynthetic
PolymeronELISA......... ........ ...... ....... ....... .. ....... ........ 59
TomozumiNoda,MasaruMatsuda,HirotakaSuzuki,
YusukeOkawa,andMotohiro Mitani
6 GasPlasmaSurfaceModificationforBiologicalAssays........ ....... ........ 73
KhorenSahagian
7 InterferenceinELISA...... ........ ....... ....... ........ ....... ........ 91
RobertS.Matson
8 DeterminationofProinflammatoryandAntiinflammatoryCytokines
byELISATechnique ....... ...... .. ...... ........ ........ ....... ........ 101
OsmanyBlancoMun˜oz
9 GyrolabImmunoassays:Miniaturization,Automation,andIntegration
intoaRapidWorkflow...... ........ ....... ....... ........ ....... ........ 109
JohanEngstr¨om
10 AssessmentofImmunologicallyPotentCarbohydrates ....... ....... ........ 129
DenongWang
11 LateralFlowMicroarray-BasedELISAforCytokines....... .. ....... ........ 141
RobertS.Matson
12 AUniqueMultiplexELISAtoProfileGrowthFactorsandCytokines
inCerebrospinalFluid...... ........ ....... ....... ........ ....... ........ 157
CharithaMadiraju,AmaniSastry,MarthaOppong,JacobKarp,
MarylaKrajewska,StanKrajewski,BarbaraTomik,AndrzejSzczudlik,
andRobertS.Matson
13 Well-BasedMultiplexFoodAllergenColorimetricELISA..... ....... ........ 169
RobertS.Matson
14 MycotoxinQuantificationbyCompetitiveELISA.... ........ ....... .... .... 183
BrynE.MerrillandRobertS.Matson
ix
x Contents
15 Lumit:AHomogeneousBioluminescentImmunoassayforDetecting
DiverseAnalytesandIntracellularProteinTargets ........... ....... ........ 195
Byounghoon(Brian)Hwang,JulianoAlves,DanLazar,NidhiNath,
LaurieEngel,MarthaO’Brien,KevinHsiao,KevinKupcho,
BeckyGodat,RodFlemming,SaidGoueli,andHichamZegzouti
16 ELISA-BasedBiosensors.... ........ ....... ....... ........ ....... ........ 225
RobertS.Matson
Index .......... ........ ....... ........ ....... ....... ........ ....... ........ 239
Contributors
JULIANOALVES • R&DDepartment,PromegaCorporation,Madison,WI,USA
BYOUNGHOON (BRIAN)HWANG • R&DDepartment,PromegaCorporation,Madison,WI,
USA;Abbvie,NorthChicago, IL,USA
LAURIEENGEL • R&DDepartment,PromegaCorporation,Madison,WI,USA
JOHANENGSTRO¨M • GyrosProteinTechnologiesAB,Uppsala,Sweden
RODFLEMMING • R&DDepartment,PromegaCorporation,Madison,WI,USA
BECKYGODAT • R&DDepartment,PromegaCorporation,Madison,WI,USA
SAIDGOUELI • R&DDepartment,PromegaCorporation,Madison,WI,USA
EMILYGRAVENS • OmegaBiologicals,Inc.,Bozeman,MT,USA
MICHAELJ.HALAT • OmegaBiologicals,Inc.,Bozeman,MT,USA
HOVHANNESHAYRAPETYAN • MarshallB.KetchumUniversity,Fullerton,CA,USA;Wayne
StateUniversity,SchoolofMedicine,Detroit,MI,USA
KEVIN HSIAO • R&DDepartment,PromegaCorporation,Madison,WI,USA
JACOBKARP • MarshallB.KetchumUniversity,Fullerton,CA,USA
MARYLAKRAJEWSKA • CellestanImmunoquant(CIQ),Inc,Oceanside,CA,USA
STANKRAJEWSKI • CellestanImmunoquant(CIQ),Inc,Oceanside,CA,USA
KEVIN KUPCHO • R&DDepartment,PromegaCorporation,Madison,WI,USA
DANLAZAR • R&DDepartment,PromegaCorporation,Madison,WI,USA
CHARITHAMADIRAJU • MarshallB.KetchumUniversity,Fullerton,CA,USA
ROBERTS.MATSON • QuantiScientificsLLC,Orange,CA,USA
MASARU MATSUDA • NOFCORPORATION,LifeScienceProductsDivision,Tokyo,Japan
BRYNE.MERRILL • ChapmanUniversity,Orange,CA,USA
MOTOHIROMITANI • NOFCORPORATION,LifeScienceProductsDivision,Tokyo,Japan
OSMANYBLANCOMUN˜OZ • MedicinayEnfermerı´a,FacultadeCienciasdelaSalud,
UniversidaddelMagdalena,SantaMarta,Magdalena,Colombia
NIDHINATH • R&DDepartment,PromegaCorporation,Madison,WI,USA;Bio-Techne,
Minneapolis, MN,USA
TOMOZUMINODA • NOFCORPORATION,LifeScienceProductsDivision,Tokyo,Japan
MARTHAO’BRIEN • R&DDepartment,PromegaCorporation,Madison,WI,USA
YUSUKEOKAWA • NOFCORPORATION,LifeScienceProductsDivision,Tokyo,Japan
MARTHAOPPONG • MarshallB.KetchumUniversity,Fullerton,CA,USA
KHORENSAHAGIAN • PlasmatreatUSAInc.,Hayward,CA,USA
AMANISASTRY • NoordaCollegeofOsteopathicMedicine,Provo,UT,USA
HIROTAKASUZUKI • NOFCORPORATION,LifeScienceProductsDivision,Tokyo,Japan
ANDRZEJSZCZUDLIK • DepartmentofNeurology,JagiellonianUniversityMedicalCollege,
Krakow,Poland
EGLIS TELLEZ-CORRALES • MarshallB.KetchumUniversity,Fullerton,CA,USA
BARBARATOMIK • DepartmentofNeurology,JagiellonianUniversityMedicalCollege,
Krakow,Poland
THAOTRAN • MarshallB.KetchumUniversity,Fullerton,CA,USA
DENONG WANG • TumorGlycomicsLaboratory,SRIInternationalBiosciencesDivision,
MenloPark,CA,USA
HICHAMZEGZOUTI • R&DDepartment,PromegaCorporation,Madison,WI,USA
xi