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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.


Related in: MedlinePlus

Dependence of the detection of Pr-DVE protein on its concentration and the well inner-surface nature ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).
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biosensors-02-00433-f004: Dependence of the detection of Pr-DVE protein on its concentration and the well inner-surface nature ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).

Mentions: The results of optical density values averaged over three experiments are shown in Figure 4. As observed before with PrPrechum, the optical density increases with the concentration of the native protein almost three-times compared to the titration run with virgin PP wells. The chemical modification of the inner-surface of the well also decreases the background noise, and therefore induces less non-specific adhesion of proteins. However, all the values of the optical density obtained with Pr-DVE are low compared with the ELISA titration of PrPrechum (Figure 1), probably due to the fact that the infectious prion protein represents only 10% of the global concentration of the native solution.


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)

Dependence of the detection of Pr-DVE protein on its concentration and the well inner-surface nature ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

biosensors-02-00433-f004: Dependence of the detection of Pr-DVE protein on its concentration and the well inner-surface nature ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).
Mentions: The results of optical density values averaged over three experiments are shown in Figure 4. As observed before with PrPrechum, the optical density increases with the concentration of the native protein almost three-times compared to the titration run with virgin PP wells. The chemical modification of the inner-surface of the well also decreases the background noise, and therefore induces less non-specific adhesion of proteins. However, all the values of the optical density obtained with Pr-DVE are low compared with the ELISA titration of PrPrechum (Figure 1), probably due to the fact that the infectious prion protein represents only 10% of the global concentration of the native solution.

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.


Related in: MedlinePlus