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Statements

Subject Item
n2:RIV%2F00027162%3A_____%2F11%3A%230000823%21RIV12-MZE-00027162
rdf:type
skos:Concept n9:Vysledek
dcterms:description
The sensitivity of antibody/hapten-based labeling systems is limited by the natural affinity ceiling of immunoglobulins. Breaking this limit by antibody engineering is difficult. We thus attempted a different approach and investigated if the so-called bridge effect, a corecognition of the linker present between hapten and carrier protein during antibody generation, can be utilized to improve the affinity of such labeling systems. The well-known haptens 2,4-dinitrophenol (2,4-DNP) and 2,4-dichlorophenoxyacetic acid (2,4-D) were equipped with various linkers, and the resulting affinity change of their cognate antibodies was analyzed by ELISA. Anti-2,4-DNP antibodies exhibited the best affinity to their hapten when it was combined with aminobutanoic acid or aminohexanoic acid. The affinity of anti-2,4-D antibodies could be enhanced even further with longer aliphatic spacers connected to the hapten. The affinity toward aminoundecanoic acid-2,4-D derivatives, for instance, was improved about 100-fold compared to 2,4-D alone and yielded detection limits as low as 100 amoles of analyte. As the effect occurred for all antibodies and haptens tested, it may be sensible to implement the bridge effect in future antibody/hapten-labeling systems in order to achieve the highest sensitivity possible. The sensitivity of antibody/hapten-based labeling systems is limited by the natural affinity ceiling of immunoglobulins. Breaking this limit by antibody engineering is difficult. We thus attempted a different approach and investigated if the so-called bridge effect, a corecognition of the linker present between hapten and carrier protein during antibody generation, can be utilized to improve the affinity of such labeling systems. The well-known haptens 2,4-dinitrophenol (2,4-DNP) and 2,4-dichlorophenoxyacetic acid (2,4-D) were equipped with various linkers, and the resulting affinity change of their cognate antibodies was analyzed by ELISA. Anti-2,4-DNP antibodies exhibited the best affinity to their hapten when it was combined with aminobutanoic acid or aminohexanoic acid. The affinity of anti-2,4-D antibodies could be enhanced even further with longer aliphatic spacers connected to the hapten. The affinity toward aminoundecanoic acid-2,4-D derivatives, for instance, was improved about 100-fold compared to 2,4-D alone and yielded detection limits as low as 100 amoles of analyte. As the effect occurred for all antibodies and haptens tested, it may be sensible to implement the bridge effect in future antibody/hapten-labeling systems in order to achieve the highest sensitivity possible.
dcterms:title
Pushing Antibody-Based Labeling Systems to Higher Sensitivity by Linker-Assisted Affinity Enhancement Pushing Antibody-Based Labeling Systems to Higher Sensitivity by Linker-Assisted Affinity Enhancement
skos:prefLabel
Pushing Antibody-Based Labeling Systems to Higher Sensitivity by Linker-Assisted Affinity Enhancement Pushing Antibody-Based Labeling Systems to Higher Sensitivity by Linker-Assisted Affinity Enhancement
skos:notation
RIV/00027162:_____/11:#0000823!RIV12-MZE-00027162
n9:predkladatel
n10:ico%3A00027162
n3:aktivita
n14:N
n3:aktivity
N
n3:cisloPeriodika
8
n3:dodaniDat
n7:2012
n3:domaciTvurceVysledku
n6:6844758
n3:druhVysledku
n13:J
n3:duvernostUdaju
n12:S
n3:entitaPredkladatele
n16:predkladatel
n3:idSjednocenehoVysledku
225294
n3:idVysledku
RIV/00027162:_____/11:#0000823
n3:jazykVysledku
n17:eng
n3:klicovaSlova
IMMUNOSORBENT ASSAYS; BRIDGE-BINDING; ANTIGEN; IMMUNOASSAYS; ELISA; RADIOIMMUNOASSAY; STREPTAVIDIN; SPECIFICITY; PERFORMANCE; DIVERSITY
n3:klicoveSlovo
n4:ANTIGEN n4:PERFORMANCE n4:RADIOIMMUNOASSAY n4:SPECIFICITY n4:STREPTAVIDIN n4:IMMUNOASSAYS n4:IMMUNOSORBENT%20ASSAYS n4:ELISA n4:BRIDGE-BINDING n4:DIVERSITY
n3:kodStatuVydavatele
US - Spojené státy americké
n3:kontrolniKodProRIV
[E92787DF78E0]
n3:nazevZdroje
BIOCONJUGATE CHEMISTRY
n3:obor
n18:CE
n3:pocetDomacichTvurcuVysledku
1
n3:pocetTvurcuVysledku
5
n3:rokUplatneniVysledku
n7:2011
n3:svazekPeriodika
22
n3:tvurceVysledku
Fránek, Milan Roeckendorf, N. Frey, A. Bade, S. Gorris, H. H.
n3:wos
000294076000019
s:issn
1043-1802
s:numberOfPages
6
n11:doi
10.1021/bc2001787