Comparison of Surface Plasmon Resonance, Resonant Waveguide Grating Biosensing and Enzyme Linked Immunosorbent Assay (ELISA) in the Evaluation of a Dengue Virus Immunoassay
Abstract
:1. Introduction
2. Material and Methods
2.1. Materials
2.2. Determination of Affinities of Anti-DENV NS1 mAbs Using a Resonant Waveguide Grating Biosensor
2.3. Kinetic Analysis of Interaction between NS1 Antigen and Antibody Using Surface Plasmon Resonance
2.4. Determination of Dengue NS1 Capture ELISA LLOD
2.5. Data Analysis
3. Results
3.1. Characterization of Anti-Dengue NS1 mAbs
Serotype origin of DENV NS1 mAbs | Number of DENV reactive mAbs characterized by immunoassays | |||
---|---|---|---|---|
DENV-1 | DENV-2 | DENV-3 | DENV-4 | |
DENV-1 | 32 | 1 | 12 | 8 |
DENV-2 | 17 | 31 | 21 | 21 |
DENV-3 | 20 | 14 | 28 | 11 |
DENV-4 | 11 | 7 | 14 | 24 |
Total | 80 | 55 | 75 | 64 |
3.2. Affinity Measurements of DENV NS1 mAbs at Saturation Using RWG
Rank of mAbs | Range of KD(M) | Number of DENV NS1 reactive mAbs in each serotype | |||
---|---|---|---|---|---|
DENV-1 | DENV-2 | DENV-3 | DENV-4 | ||
High-affinity | 10−8 | 8 | 19 | 12 | 9 |
Moderate-affinity | 10−7 | 29 | 14 | 22 | 14 |
Low-affinity | 10−6 | 29 | 17 | 21 | 26 |
10−5 | 9 | 2 | 14 | 14 | |
10−4 | 1 | 0 | 1 | 0 | |
Non-binders | <10−3* | 4 | 3 | 5 | 1 |
Total | 80 | 55 | 75 | 64 |
3.3. Binding Kinetics of NS1 mAbs Using SPR
3.4. Affinity Comparison of NS1 mAbs between RWG and SPR
Dengue NS1 serotype | LLOD vs. RWG KD | LLOD vs. SPR KD | ||
---|---|---|---|---|
Correlation co-efficient (r2) | P value | Correlation co-efficient (r2) | P value | |
DENV-1 | 0.5506 | 0.0024 | 0.008603 | 0.8124 |
DENV-2 | 0.2804 | 0.2216 | 0.1409 | 0.4068 |
DENV-3 | 0.576 | 0.0479 | 0.3676 | 0.3937 |
DENV-4 | 0.07368 | 0.7286 | 0.03127 | 0.8232 |
3.5. Correlation of SPR and RWG Affinity with Capture ELISA LLOD
MAb ID | Serotype specificity | LLOD (pg/mL) | RWG KD (nM) | SPR KD (nM) |
---|---|---|---|---|
Dv1 M32 | 1 | 4.1 | 44 | 37 |
Dv1 M19 | 1 | 4.2 | 103 | 36 |
Dv1 M1 | 1 | 4.3 | 25 | 37 |
Dv3 M7 | 1 | 4.5 | 54 | 7 |
Gus 2 | 1 | 4.7 | 87 | 12 |
Dv3 M18 | 1 | 4.9 | 414 | 27 |
Dv2 M27 | 1 | 5.6 | 137 | 83 |
Dv3 M4 | 1 | 6.8 | 99 | 61 |
Dv1 M33 | 1 | 3.0 | 296 | 11 |
Dv2 M4 | 2 | 3.0 | 77 | 23 |
Dv2 M27 | 2 | 3.2 | 28 | 7 |
Dv2 M10 | 2 | 3.2 | 60 | 12 |
Gus 2 | 2 | 3.2 | 75 | 7 |
Dv2 M17 | 2 | 3.4 | 101 | 38 |
Dv2 M14 | 2 | 3.8 | 124 | 16 |
Dv3 M4 | 2 | 4.1 | 94 | 27 |
Gus2 | 3 | 4.7 | 96 | 42 |
Dv3 M11 | 3 | 4.7 | 86 | 10 |
Dv3 M7 | 3 | 5.4 | 44 | 5 |
Dv3 M4 | 3 | 4.2 | 105 | 11 |
Dv4 M27 | 4 | 4.2 | 506 | 141 |
Dv3 M7 | 4 | 4.4 | 47 | 12 |
Dv4 M6 | 4 | 5.0 | 134 | 33 |
Gus 2 | 4 | 5.5 | 99 | 31 |
Dv3 M4 | 4 | 4.1 | 99 | 38 |
4. Conclusion and Discussion
Acknowledgments
Conflict of Interest
References
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Hu, D.; Fry, S.R.; Huang, J.X.; Ding, X.; Qiu, L.; Pan, Y.; Chen, Y.; Jin, J.; McElnea, C.; Buechler, J.; et al. Comparison of Surface Plasmon Resonance, Resonant Waveguide Grating Biosensing and Enzyme Linked Immunosorbent Assay (ELISA) in the Evaluation of a Dengue Virus Immunoassay. Biosensors 2013, 3, 297-311. https://doi.org/10.3390/bios3030297
Hu D, Fry SR, Huang JX, Ding X, Qiu L, Pan Y, Chen Y, Jin J, McElnea C, Buechler J, et al. Comparison of Surface Plasmon Resonance, Resonant Waveguide Grating Biosensing and Enzyme Linked Immunosorbent Assay (ELISA) in the Evaluation of a Dengue Virus Immunoassay. Biosensors. 2013; 3(3):297-311. https://doi.org/10.3390/bios3030297
Chicago/Turabian StyleHu, Dongmei, Scott R. Fry, Johnny X. Huang, Xixia Ding, Liwen Qiu, Yuxian Pan, Yue Chen, Jing Jin, Catriona McElnea, Joe Buechler, and et al. 2013. "Comparison of Surface Plasmon Resonance, Resonant Waveguide Grating Biosensing and Enzyme Linked Immunosorbent Assay (ELISA) in the Evaluation of a Dengue Virus Immunoassay" Biosensors 3, no. 3: 297-311. https://doi.org/10.3390/bios3030297