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.


Related in: MedlinePlus

Dependence of the (D – D0)/D0 ratio on the well inner-surface nature and the antigen concentration. ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).
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biosensors-02-00433-f002: Dependence of the (D – D0)/D0 ratio on the well inner-surface nature and the antigen concentration. ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).

Mentions: In Figure 2, the results are presented through a normalization step ((OD of measured sample-OD of control PP)/OD of control PP) corresponding to a sensitivity of the biosensor. Figure 2 confirms clearly that the treated substrates exhibit greater susceptibility to prion protein than the control, especially with T1 and T2 functionalization. The presence of a surface charge onto PP would therefore better fix the capture antibody by electrostatic effects and van der Waals interactions, and thus better detect the protein. From Figure 2, the detection threshold of PrPrechum is determined as 25 ng∙mL–1, 9.4 ng∙mL–1, 9.3 ng∙mL–1 and 14.2 ng∙mL–1 for virgin PP, PP-T1, PP-T2 and PP-T3, respectively. Indeed, the sensitivity is enhanced with T1 and T2 treatments, while T3 treatment has no significant improvement compared to virgin support, then it would confirm the need for a surface charge effect. The limit of detection (%∙ng–1∙mL–1) corresponding to the cross point between the slopes of insensitivity and sensitivity curves is also more important when T1 and T2 surface modifications are applied onto PP well: 47.9 for PP-T2 > 36.8 for PP-T1 > 22.5 for virgin PP > 19.8 for PP-T3. Compared to the results obtained with neuroproteins involved in Parkinson’s disease, it shows that the capture antibody has no affinity for the PP-T1 substrate. Thus, the sensitivity of the PP-T1 substrate in ELISA-assays is expected to be low. In contrast to that, the sensitivity of the PP-T1 substrate in the ELISA assays in the present work is as high as the one measured on the PP-T2 and PP-T3 substrates. This fact could be explained by different surface (physico-chemical or conformation) properties of both types of proteins in various biological media.


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 (D – D0)/D0 ratio on the well inner-surface nature and the antigen concentration. ((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-f002: Dependence of the (D – D0)/D0 ratio on the well inner-surface nature and the antigen concentration. ((capture antibody) = 10 µg∙mL–1, (detection antibody) = 1 µg∙mL–1).
Mentions: In Figure 2, the results are presented through a normalization step ((OD of measured sample-OD of control PP)/OD of control PP) corresponding to a sensitivity of the biosensor. Figure 2 confirms clearly that the treated substrates exhibit greater susceptibility to prion protein than the control, especially with T1 and T2 functionalization. The presence of a surface charge onto PP would therefore better fix the capture antibody by electrostatic effects and van der Waals interactions, and thus better detect the protein. From Figure 2, the detection threshold of PrPrechum is determined as 25 ng∙mL–1, 9.4 ng∙mL–1, 9.3 ng∙mL–1 and 14.2 ng∙mL–1 for virgin PP, PP-T1, PP-T2 and PP-T3, respectively. Indeed, the sensitivity is enhanced with T1 and T2 treatments, while T3 treatment has no significant improvement compared to virgin support, then it would confirm the need for a surface charge effect. The limit of detection (%∙ng–1∙mL–1) corresponding to the cross point between the slopes of insensitivity and sensitivity curves is also more important when T1 and T2 surface modifications are applied onto PP well: 47.9 for PP-T2 > 36.8 for PP-T1 > 22.5 for virgin PP > 19.8 for PP-T3. Compared to the results obtained with neuroproteins involved in Parkinson’s disease, it shows that the capture antibody has no affinity for the PP-T1 substrate. Thus, the sensitivity of the PP-T1 substrate in ELISA-assays is expected to be low. In contrast to that, the sensitivity of the PP-T1 substrate in the ELISA assays in the present work is as high as the one measured on the PP-T2 and PP-T3 substrates. This fact could be explained by different surface (physico-chemical or conformation) properties of both types of proteins in various biological media.

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