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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

Comparison of SPR determined percent activity of mAbs betweenm Abs of different affinity subset, 10 nM and 100 nM in RWG. P value above the dots represents the statistical difference between both subsets of mAbs. In this study, except DENV-4 reactive mAbs, the percent activities of the other three mAb serotypes were significantly higher in mAbs with affinity of 10 nM (RWG) than that of with affinity of 100nM (RWG) using Mann-Whitney test.
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biosensors-03-00297-f005: Comparison of SPR determined percent activity of mAbs betweenm Abs of different affinity subset, 10 nM and 100 nM in RWG. P value above the dots represents the statistical difference between both subsets of mAbs. In this study, except DENV-4 reactive mAbs, the percent activities of the other three mAb serotypes were significantly higher in mAbs with affinity of 10 nM (RWG) than that of with affinity of 100nM (RWG) using Mann-Whitney test.

Mentions: For mAbs where affinities could be determined accurately by RWG and SPR, correlation analysis using the Spearman test demonstrated that DENV-1 (R = −0.1868, P = 0.3507) and DENV-3 (R = −0.05147, P = 0.8445) NS1 reactive mAbs RWG and SPR affinities were not correlated. However, DENV-2 (R = 0.3656, P = 0.0394) and DENV-4 (R = 0.7559, P = 0.0007) NS1 reactive mAbs displayed a moderate correlation. The percentage activities of the antibodies (the amount determined by comparison of the theoretical and experimental maximal response; see Section 2.5) determined by SPR and RWG were correlated within different affinity subsets. The Mann-Whitney test demonstrated that the percentage activities of DENV-1 (U = 10, P = 0.0002), DENV-2 (U = 47, P = 0.0003) and DENV-3 (U = 9, P = 0.0182) NS1 reactive mAbs were significantly higher in the RWG high-affinity mAbs subsets than that of RWG middle-affinity mAbs. Although the difference of percentage activity of DENV-4 reactive mAbs was not so significant (U = 18, P = 0.1738), they displayed the same trend (Figure 5).


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)

Comparison of SPR determined percent activity of mAbs betweenm Abs of different affinity subset, 10 nM and 100 nM in RWG. P value above the dots represents the statistical difference between both subsets of mAbs. In this study, except DENV-4 reactive mAbs, the percent activities of the other three mAb serotypes were significantly higher in mAbs with affinity of 10 nM (RWG) than that of with affinity of 100nM (RWG) using Mann-Whitney test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

biosensors-03-00297-f005: Comparison of SPR determined percent activity of mAbs betweenm Abs of different affinity subset, 10 nM and 100 nM in RWG. P value above the dots represents the statistical difference between both subsets of mAbs. In this study, except DENV-4 reactive mAbs, the percent activities of the other three mAb serotypes were significantly higher in mAbs with affinity of 10 nM (RWG) than that of with affinity of 100nM (RWG) using Mann-Whitney test.
Mentions: For mAbs where affinities could be determined accurately by RWG and SPR, correlation analysis using the Spearman test demonstrated that DENV-1 (R = −0.1868, P = 0.3507) and DENV-3 (R = −0.05147, P = 0.8445) NS1 reactive mAbs RWG and SPR affinities were not correlated. However, DENV-2 (R = 0.3656, P = 0.0394) and DENV-4 (R = 0.7559, P = 0.0007) NS1 reactive mAbs displayed a moderate correlation. The percentage activities of the antibodies (the amount determined by comparison of the theoretical and experimental maximal response; see Section 2.5) determined by SPR and RWG were correlated within different affinity subsets. The Mann-Whitney test demonstrated that the percentage activities of DENV-1 (U = 10, P = 0.0002), DENV-2 (U = 47, P = 0.0003) and DENV-3 (U = 9, P = 0.0182) NS1 reactive mAbs were significantly higher in the RWG high-affinity mAbs subsets than that of RWG middle-affinity mAbs. Although the difference of percentage activity of DENV-4 reactive mAbs was not so significant (U = 18, P = 0.1738), they displayed the same trend (Figure 5).

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