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Comparison of surface plasmon resonance, resonant waveguide grating biosensing and enzyme linked immunosorbent assay (ELISA) in the evaluation of a dengue virus immunoassay.

Hu D, Fry SR, Huang JX, Ding X, Qiu L, Pan Y, Chen Y, Jin J, McElnea C, Buechler J, Che X, Cooper MA - Biosensors (Basel) (2013)

Bottom Line: However, there is a significant scope to improve both the sensitivity and the specificity of those tests.The interactions of antibody (Ab)-antigen (Ag) were profiled, with weak interactions (KD = 1-0.1 μM) able to be detected under static equilibrium conditions by RWG, but not observed to under more rigorous flow conditions using SPR.Hence, whilst high-throughput RWG can be useful as preliminary screening for higher affinity antibodies, care should be exercised in the assignation of quantitative values for affinity between different assay formats.

View Article: PubMed Central - PubMed

Affiliation: Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, University of Queensland, Brisbane, 4072 Australia. zjyyhdm@163.com.

ABSTRACT
Two label-free biosensor platforms, Resonance Waveguide Grating (RWG) and Surface Plasmon Resonance (SPR), were used to rank a large panel of anti-dengue virus NS1 antibodies. Dengue non-structural 1 (NS1) protein is an established serological marker for the early detection of dengue infection. A variety of commercial dengue NS1 antigen capture immunoassays are available in both ELISA and lateral flow format. However, there is a significant scope to improve both the sensitivity and the specificity of those tests. The interactions of antibody (Ab)-antigen (Ag) were profiled, with weak interactions (KD = 1-0.1 μM) able to be detected under static equilibrium conditions by RWG, but not observed to under more rigorous flow conditions using SPR. There were significant differences in the absolute affinities determined by the two technologies, and there was a poor correlation between antibodies best ranked by RWG and the lower limit of detection (LLOD) found by ELISA. Hence, whilst high-throughput RWG can be useful as preliminary screening for higher affinity antibodies, care should be exercised in the assignation of quantitative values for affinity between different assay formats.

No MeSH data available.


Related in: MedlinePlus

Representative binding isotherms for mAbs binding to non-structural 1 (NS1) measured by Resonance Waveguide Grating (RWG). (A) shows the saturated binding model of NS1 mAb (Dv1M1) to immobilized DENV NS1 protein, which generally produced middle to high affinity in RWG; (B) demonstrates the non-saturable binding model (Dv1 M4), which generally corresponded with low affinity. Values were given as mean ± standard error of the mean for n = 4.
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biosensors-03-00297-f001: Representative binding isotherms for mAbs binding to non-structural 1 (NS1) measured by Resonance Waveguide Grating (RWG). (A) shows the saturated binding model of NS1 mAb (Dv1M1) to immobilized DENV NS1 protein, which generally produced middle to high affinity in RWG; (B) demonstrates the non-saturable binding model (Dv1 M4), which generally corresponded with low affinity. Values were given as mean ± standard error of the mean for n = 4.

Mentions: RWG response values (picometer change in signal) of each mAb were plotted against the serial concentrations using GraphPad Prism® software. The KD values were calculated using a non-linear regression model as described in the methods section (Figure 1). On the basis of the plots, the calculated affinity, KD values of these mAbs were determined (Figure 2) with a wide spectrum of affinity spanning nM to mM. In this study, mAbs with KD ≥ 1 μM were determined as low affinity mAbs and not studied further (Table 2).


Comparison of surface plasmon resonance, resonant waveguide grating biosensing and enzyme linked immunosorbent assay (ELISA) in the evaluation of a dengue virus immunoassay.

Hu D, Fry SR, Huang JX, Ding X, Qiu L, Pan Y, Chen Y, Jin J, McElnea C, Buechler J, Che X, Cooper MA - Biosensors (Basel) (2013)

Representative binding isotherms for mAbs binding to non-structural 1 (NS1) measured by Resonance Waveguide Grating (RWG). (A) shows the saturated binding model of NS1 mAb (Dv1M1) to immobilized DENV NS1 protein, which generally produced middle to high affinity in RWG; (B) demonstrates the non-saturable binding model (Dv1 M4), which generally corresponded with low affinity. Values were given as mean ± standard error of the mean for n = 4.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263579&req=5

biosensors-03-00297-f001: Representative binding isotherms for mAbs binding to non-structural 1 (NS1) measured by Resonance Waveguide Grating (RWG). (A) shows the saturated binding model of NS1 mAb (Dv1M1) to immobilized DENV NS1 protein, which generally produced middle to high affinity in RWG; (B) demonstrates the non-saturable binding model (Dv1 M4), which generally corresponded with low affinity. Values were given as mean ± standard error of the mean for n = 4.
Mentions: RWG response values (picometer change in signal) of each mAb were plotted against the serial concentrations using GraphPad Prism® software. The KD values were calculated using a non-linear regression model as described in the methods section (Figure 1). On the basis of the plots, the calculated affinity, KD values of these mAbs were determined (Figure 2) with a wide spectrum of affinity spanning nM to mM. In this study, mAbs with KD ≥ 1 μM were determined as low affinity mAbs and not studied further (Table 2).

Bottom Line: However, there is a significant scope to improve both the sensitivity and the specificity of those tests.The interactions of antibody (Ab)-antigen (Ag) were profiled, with weak interactions (KD = 1-0.1 μM) able to be detected under static equilibrium conditions by RWG, but not observed to under more rigorous flow conditions using SPR.Hence, whilst high-throughput RWG can be useful as preliminary screening for higher affinity antibodies, care should be exercised in the assignation of quantitative values for affinity between different assay formats.

View Article: PubMed Central - PubMed

Affiliation: Division of Chemistry and Structural Biology, Institute for Molecular Bioscience, University of Queensland, Brisbane, 4072 Australia. zjyyhdm@163.com.

ABSTRACT
Two label-free biosensor platforms, Resonance Waveguide Grating (RWG) and Surface Plasmon Resonance (SPR), were used to rank a large panel of anti-dengue virus NS1 antibodies. Dengue non-structural 1 (NS1) protein is an established serological marker for the early detection of dengue infection. A variety of commercial dengue NS1 antigen capture immunoassays are available in both ELISA and lateral flow format. However, there is a significant scope to improve both the sensitivity and the specificity of those tests. The interactions of antibody (Ab)-antigen (Ag) were profiled, with weak interactions (KD = 1-0.1 μM) able to be detected under static equilibrium conditions by RWG, but not observed to under more rigorous flow conditions using SPR. There were significant differences in the absolute affinities determined by the two technologies, and there was a poor correlation between antibodies best ranked by RWG and the lower limit of detection (LLOD) found by ELISA. Hence, whilst high-throughput RWG can be useful as preliminary screening for higher affinity antibodies, care should be exercised in the assignation of quantitative values for affinity between different assay formats.

No MeSH data available.


Related in: MedlinePlus