Limits...
A New Approach for Detection Improvement of the Creutzfeldt-Jakob Disorder through a Specific Surface Chemistry Applied onto Titration Well.

Mille C, Debarnot D, Zorzi W, Moualij BE, Quadrio I, Perret-Liaudet A, Coudreuse A, Legeay G, Poncin-Epaillard F - Biosensors (Basel) (2012)

Bottom Line: It is achieved thanks to the association of plasma chemistry and coating with different amphiphilic molecules bearing either ionic charges and/or long hydrocarbon chains.The treated support by 3-butenylamine hydrochloride improves the signal detection of recombinant protein, while surface modification with the 3,7-dimethylocta-2,6-dien-1-diamine (geranylamine) enhances the sensitivity of the native protein.Beside the surface chemistry effect, these different results are associated with protein conformation.

View Article: PubMed Central - PubMed

Affiliation: Université, UMR Université du Maine, CNRS n°6283, Institut des Molécules et Matériaux du Mans, Département Polymères, Colloïdes et Interfaces, av. O. Messiaen, 72085 Le Mans, France. caroline.mille.etu@univ-lemans.fr.

ABSTRACT
This work illustrates the enhancement of the sensitivity of the ELISA titration for recombinant human and native prion proteins, while reducing other non-specific adsorptions that could increase the background signal and lead to a low sensitivity and false positives. It is achieved thanks to the association of plasma chemistry and coating with different amphiphilic molecules bearing either ionic charges and/or long hydrocarbon chains. The treated support by 3-butenylamine hydrochloride improves the signal detection of recombinant protein, while surface modification with the 3,7-dimethylocta-2,6-dien-1-diamine (geranylamine) enhances the sensitivity of the native protein. Beside the surface chemistry effect, these different results are associated with protein conformation.

No MeSH data available.


Evaluation of non-specific associations for each surface treatment during Pr-DVE ELISA titration.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4263562&req=5

biosensors-02-00433-f007: Evaluation of non-specific associations for each surface treatment during Pr-DVE ELISA titration.

Mentions: In Figure 7, the variations of obtained optical density values based on the affinity of each biomolecule for each relative support treatment correspond to the Pr-DVE titration. The virgin PP well seems to fix the antigen and/or detection antibody in absence of the capture. Indeed, the value of the associated optical density is two to eight times greater than those of treated substrates. Compared to the other treatments, PP-T1 coated with bovine serum albumin (BSA) as a blocking agent gives rise to non-specific binding of the antigen and/or detection antibody. In absence of the antigen, the measured optical densities are low—below 0.1 a.u.—and of half value for the treated supports compared to the virgin well. This indicates a low detection antibody adsorption onto the capture antibody and/or the blocking agent. The presence of charges and amine groups on the surface of polypropylene therefore reduces background noise on the binding of secondary antibody to primary antibody. Moreover, the last experiment, corresponding to the absence of detection antibody, confirms that the weak obtained signal is due to the association of antibodies with each other. The T2 surface chemistry seems to be the most appropriate to the Pr-DVE detection because of the low optical densities in each experiment (below 0.1 a.u.), and therefore it decreases the background noise associated to non-specific binding.


A New Approach for Detection Improvement of the Creutzfeldt-Jakob Disorder through a Specific Surface Chemistry Applied onto Titration Well.

Mille C, Debarnot D, Zorzi W, Moualij BE, Quadrio I, Perret-Liaudet A, Coudreuse A, Legeay G, Poncin-Epaillard F - Biosensors (Basel) (2012)

Evaluation of non-specific associations for each surface treatment during Pr-DVE ELISA titration.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263562&req=5

biosensors-02-00433-f007: Evaluation of non-specific associations for each surface treatment during Pr-DVE ELISA titration.
Mentions: In Figure 7, the variations of obtained optical density values based on the affinity of each biomolecule for each relative support treatment correspond to the Pr-DVE titration. The virgin PP well seems to fix the antigen and/or detection antibody in absence of the capture. Indeed, the value of the associated optical density is two to eight times greater than those of treated substrates. Compared to the other treatments, PP-T1 coated with bovine serum albumin (BSA) as a blocking agent gives rise to non-specific binding of the antigen and/or detection antibody. In absence of the antigen, the measured optical densities are low—below 0.1 a.u.—and of half value for the treated supports compared to the virgin well. This indicates a low detection antibody adsorption onto the capture antibody and/or the blocking agent. The presence of charges and amine groups on the surface of polypropylene therefore reduces background noise on the binding of secondary antibody to primary antibody. Moreover, the last experiment, corresponding to the absence of detection antibody, confirms that the weak obtained signal is due to the association of antibodies with each other. The T2 surface chemistry seems to be the most appropriate to the Pr-DVE detection because of the low optical densities in each experiment (below 0.1 a.u.), and therefore it decreases the background noise associated to non-specific binding.

Bottom Line: It is achieved thanks to the association of plasma chemistry and coating with different amphiphilic molecules bearing either ionic charges and/or long hydrocarbon chains.The treated support by 3-butenylamine hydrochloride improves the signal detection of recombinant protein, while surface modification with the 3,7-dimethylocta-2,6-dien-1-diamine (geranylamine) enhances the sensitivity of the native protein.Beside the surface chemistry effect, these different results are associated with protein conformation.

View Article: PubMed Central - PubMed

Affiliation: Université, UMR Université du Maine, CNRS n°6283, Institut des Molécules et Matériaux du Mans, Département Polymères, Colloïdes et Interfaces, av. O. Messiaen, 72085 Le Mans, France. caroline.mille.etu@univ-lemans.fr.

ABSTRACT
This work illustrates the enhancement of the sensitivity of the ELISA titration for recombinant human and native prion proteins, while reducing other non-specific adsorptions that could increase the background signal and lead to a low sensitivity and false positives. It is achieved thanks to the association of plasma chemistry and coating with different amphiphilic molecules bearing either ionic charges and/or long hydrocarbon chains. The treated support by 3-butenylamine hydrochloride improves the signal detection of recombinant protein, while surface modification with the 3,7-dimethylocta-2,6-dien-1-diamine (geranylamine) enhances the sensitivity of the native protein. Beside the surface chemistry effect, these different results are associated with protein conformation.

No MeSH data available.