Development of EndoScreen Chip, a Microfluidic Pre-Endoscopy Triage Test for Esophageal Adenocarcinoma
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Study Cohort and Experimental Design
2.2. Recombinant Protein Expression and Purification
2.3. Antibody Production and Purification
2.4. Serum Purified C9 and Production of Depleted Serum
2.5. Characterization of mAb 26 by Western Blot
2.6. Hemolytic Assay
2.7. C9 Direct ELISA
2.8. EndoScreen Chip Fabrication and Functionalization
2.9. SERS Nanotags Synthesis
2.10. C9 Sample Preparation for EndoScreen Chip Assay
2.11. EndoScreen Chip Assay
2.12. Statistical Analysis
3. Results
3.1. Validation of New C9 Monoclonal Antibody
3.2. Establishing the Endo Screen Chip
3.3. Total C9 and JAC-C9 Are Elevated in EAC Patient Serum
3.4. Serum C9 Biomarker Panel Improves the Detection of EAC over Clinical Risk Factors Alone
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Parameter | Category | Healthy | BE | EAC | p Value (Healthy vs. BE vs. EAC) |
---|---|---|---|---|---|
Number | 15 | 16 | 15 | ||
Age (years) | Median | 64.39 | 61.89 | 61.72 | 0.146 a |
Range | 56–75 | 52–75 | 53–74 | ||
BMI | Healthy wt (<25) | 10.9% (5) | 6.5% (3) | 2.2% (1) | 0.054 b |
Overweight (<30) | 10.9% (5) | 23.9% (11) | 10.9% (5) | ||
Obese I (<35) | 10.9% (5) | 4.3% (2) | 17.4% (8) | ||
Obese II (<40) | 0.0% (0) | 0.0% (0) | 0.0% (0) | ||
Obese III (≥40) | 0.0% (0) | 0.0% (0) | 2.2% (1) | ||
Heartburn and Reflux History | Never | 11.1% (5) | 4.4% (2) | 2.2% (1) | 0.005 b |
<Once/month | 6.7% (3) | 6.7% (3) | 2.2% (1) | ||
Monthly (few times/month) | 8.9% (4) | 13.3% (6) | 2.2% (1) | ||
Weekly (few times/wk) | 2.2% (1) | 0.0% (0) | 15.6% (7) | ||
Daily | 2.2% (1) | 11.1% (5) | 11.1% (5) |
Predictive model | Health Status: Healthy | ||||
---|---|---|---|---|---|
Predictor | AUC | StdError | Lower 95% | Upper 95% | Prob > ChiSq |
Clinical Risk Model | 0.7350 | 0.0886 | 0.5322 | 0.8712 | |
Biomarker Model | 0.8272 | 0.0621 | 0.6714 | 0.9181 | |
Difference | −0.092 | 0.0635 | −0.217 | 0.0323 | 0.1465 |
Health Status: BE | |||||
Predictor | AUC | StdError | Lower 95% | Upper 95% | Prob > ChiSq |
Clinical Risk Model | 0.7435 | 0.0726 | 0.5788 | 0.8595 | |
Biomarker Model | 0.8405 | 0.0579 | 0.6934 | 0.9247 | |
Difference | −0.097 | 0.0690 | −0.232 | 0.0382 | 0.1598 |
Health Status: EAC | |||||
Predictor | AUC | StdError | Lower 95% | Upper 95% | Prob > ChiSq |
Clinical Risk Model | 0.8378 | 0.0624 | 0.6774 | 0.9270 | |
Biomarker Model | 0.9311 | 0.0411 | 0.7936 | 0.9794 | |
Difference | −0.093 | 0.0432 | −0.178 | −0.009 | 0.0309 * |
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Webster, J.A.; Wuethrich, A.; Shanmugasundaram, K.B.; Richards, R.S.; Zelek, W.M.; Shah, A.K.; Gordon, L.G.; Kendall, B.J.; Hartel, G.; Morgan, B.P.; et al. Development of EndoScreen Chip, a Microfluidic Pre-Endoscopy Triage Test for Esophageal Adenocarcinoma. Cancers 2021, 13, 2865. https://doi.org/10.3390/cancers13122865
Webster JA, Wuethrich A, Shanmugasundaram KB, Richards RS, Zelek WM, Shah AK, Gordon LG, Kendall BJ, Hartel G, Morgan BP, et al. Development of EndoScreen Chip, a Microfluidic Pre-Endoscopy Triage Test for Esophageal Adenocarcinoma. Cancers. 2021; 13(12):2865. https://doi.org/10.3390/cancers13122865
Chicago/Turabian StyleWebster, Julie A., Alain Wuethrich, Karthik B. Shanmugasundaram, Renee S. Richards, Wioleta M. Zelek, Alok K. Shah, Louisa G. Gordon, Bradley J. Kendall, Gunter Hartel, B. Paul Morgan, and et al. 2021. "Development of EndoScreen Chip, a Microfluidic Pre-Endoscopy Triage Test for Esophageal Adenocarcinoma" Cancers 13, no. 12: 2865. https://doi.org/10.3390/cancers13122865