简介：Studyonbiotinylationstrategiesforcompetitiveimmunoassayofestradiol(E2)wascarriedout.Twotypesofcompetitiveenzymeimmunoassay(EIA)withBiotin-Avidinamplificationsystemwereestablishedandoptimized.TheE2-Biotinconjugatewasusedasatracerinoneassay,andbiotinylatedantibodywasusedasatracerintheother.InbothofEIAs,horseradishperoxidase-labelledAvidin(Avidin-HRP)wasusedwithaspectrometricdeterminationofenzymeactivity.Theprecision,sensitivityandspecificityweremeasuredandcompared.Theresultsshowedthatalthoughbothweresatisfactoryinspecificity,theEIAwithhapten-BiotinshowedtobesuperiortotheEIAwithbiotinylatedantibodyinsensitivityandprecision.ThelimitofdetectionofserumE2was8and50pg/mLwithE2-Biotinandbiotinylatedantibodyastracer,respectively.

简介：Objective:ToevaluatethevalueofenhancedluminescenceenzymeimmunoassayinthediagnosisofNeisseriagonorrhea(NG)infection.Methods:Anti-catalaseantibodyforNeisseriagonorrheaecombinedwithenhancedluminescenceenzymeimmunoassaywereusedtotestforN.gonorrhea.Results:Aminimumof1×10^4/CFUofGCingenitaltractsecretionsorurinecouldbedetectedwiththetechniqueofluminescenceenzymeimmunoassay.Conclusion:TheenhancedluminescenceenzymeimmunoassayhastheadvantageofhighsensitivityandspecificityfordiagnosingNGfromgenitourinarytractsecretionandurine.

简介：AnewchemilurninescencelabelN-(β-carboxypropionyl)luminol(CPL)wasusedtolabelsheepanti-humanIgG(SaHIgG).Thelabeledantibodywasstableandcouldbedetectedatleastdownto10-17~10-16mol.Themolarincorporationratiowasestimatedtobe0.26molofCPLpermolofSaHIgG.TherewerenoapparentchangesintheimmunoreactivityofthelabeledSaHIgGandinthequantumefficiencyoftheCPLafterlabeling.

简介：Preparationandcharacterizationofthehapten-proteinconjugatesarefundamentaltodevelopingenvironmentalimmunoassays.Asahapten,1-pyrenebutyricacid(PBA)wasconjugatedtothecarrierproteinofbovineserumalbumin(BSA)orovalbumin(OVA)byactiveestermethod.Infraredspectra(IR)showedthatPBA-BSAandPBA-OVAconjugatesweresuccessfullyprepared.Thenumberofthehaptensconjugatedtothecarrierproteinwasdeterminedbyultravioletspectra(UV)andmatrix-assistedlaserdesorptionionizationtime-of-flightmassspectrometry(MALDI-TOF-MS).ThecalculatedaveragebindingratiosofPBA/BSAandPBA/OVAwere18:1and10:1byUV,and31:1and22:1byMALDI-TOF-MS,respectively.Althoughtherewasadiscrepancybetweentheresultsdeterminedbythetwomethods,bothofthemwereusefulforthecharacterizationofthehapten-proteinconjugates.TheantibodywasproducedagainsttheantigenofPBA-BSA,andtheaffinitywastestedbythedoubleagardiffusionmethod.Theconjugatesandtheantibodycouldbeusedfordevelopingasensitiveandselectiveimmunoassayofpolycyclicaromatichydrocarbons(PAHs).

简介：Inthispaper,itwasdiscoveredthatanovelpH-sensitivecopolymerofN-isopropylacrylamide(NIP)andN-(3-dimethylaminopropyl)methacrylamide(DMAPM)couldbegottenbypolymerization.ThephasetransitionpH(pHtr)ofP(NIP-DMAPM)polymerwasfoundtobe7.4at37℃.ThepolymerwasprecipitatedoutofwateraboveacriticalpH=7.4andre-dissolvedbelowpH----7.4.Thecharacteristicofthispolymermadeitpossibletocarryouttheimmunochemicalstepsofanimmunoassayinatruesolutionandthentoquicklyseparatetheresultingproductfromthereactionmixture.Inacompetitivefluorescenceimmunoassay,thestandardrabbitIgGandrabbitIgGimmobilizedonP(NIP-DMAPM)firstcompetitivelyreactedwiththefluoresceinisothiocyanate(FITC)labeledantibody,thenthepHofsolutionwasadjustedabovethepHtrofpolymertoprecipitatethepolymer-immunecomplex,andthepolymer-immunecomplexprecipitatewasseparatedandre-dissolvedbytheadjustmentofpH,finallytheFITC-labeledantibodyintheimmunecomplexwasquantifiedbyfluorescencemeasurement.ThecalibrationgraphforrabbitIgGwaslinearovertherangeof100-1000ng/mLwithadetectionlimitof11ng/mL.Themethodisrapid,sensitiveandsimple.OwingtoneutralpHtrofP(NIP-DMAPM),thedamagetoantigen-antibodyimmunecomplexwasgreatlydecreasedinthecourseofseparation.Inaddition,asandwichenzyme-linkedfluorescenceimmunoassaymethodforthedeterminationofhumanIgGwasalsodeveloped,showingthatthepH-sensitivephaseseparatingimmunoassaycouldbeperformedinthecompetitivemethodaswellasthesandwichmethod.

简介：AbstractLike antibody evaluation, using an effective antigen-specific T-cell immunity assessment method in coronavirus disease 2019 (COVID-19) patients, survivors and vaccinees is crucial for understanding the immune persistence, prognosis assessment, and vaccine development for COVID-19. This study evaluated an empirically adjusted enzyme-linked immunospot assay for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific T-cell immunity in 175 peripheral blood samples from COVID-19 convalescents and healthy individuals. Results of viral nucleic acid were used as the gold standard of infection confirmation. The SARS-CoV-2M peptide pool had higher sensitivity of 85% and specificity of 71% for the single peptide pool. For combined peptide pools, the parallel evaluation (at least one of the peptide pools is positive) of total peptide pools (S1&S2&M&N) had higher sensitivity (up to 93%), and the serial evaluation (all peptide pools are positive) of total peptide pools had higher specificity (up to 100%). The result of the serial evaluation was better than that of the parallel evaluation as a whole. The detection efficiency of M and N peptide pool serial evaluation appeared the highest, with a sensitivity of 80% and specificity of 93%. This T-cell immunity detection assay introduced in this report can achieve high operability and applicability. Therefore, it can be an effective SARS-CoV-2-specific cellular immune function evaluation method.