Research biosensor

1. CENTER OF BIOMOLECUL AR THERAPY AND DIAGNOSTICS (CBTD);
CENTER OF INNOVATIVE IMMUNODIAGNOSTIC DEVELOPMENT (CIID)
FACULTY OF ASSOCIATED MEDICAL SCIENCES, CHIANG MAI UNIVERSITY
13 January 2022
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Chatchai Tayapiwatana
Point-of-Care test kit for screening of
anti-interferon-gamma antibody in patients with
adult-onset immunodeficiency (AOID)

2. Objective
- To establish a collaboration with expert clinician in AOID
- To validate the IC strip with clinical samples
- To opitimize the sensitivity & specificity of IC strip
- To identify the application of IC strip (screening or monitoring)
- To understand current diagnostic and therapeutic situation
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3. Introduction : Interferon gamma (IFN-γ)
https://commons.wikimedia.org/wiki/File:Interferon_Gamma.png
• Dimerized soluble cytokine
• Type II Interferon
• Antiviral activity
• Produced predominantly by
• Natural killer cells (NK cells)
• T-cells
• Monomer consists of
• 6 α-helices
• 143 amino acids
• Receptors: IFNGR1 and IFNGR2
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4. 4
Adult-onset immunodeficiency syndrome (AOID)
• A newly diagnosed Immunodeficiency illness, PID
• The mechanism of disease remains obscure
• Genetic defect
• Affecting mainly people of Asian aged 40-50
• Cases first started appearing in 2004
 primarily in Thailand and Taiwan
• Patients produce autoantibodies against
interferon-gamma (Anti IFN-γ)

5. Patients with adult-onset immunodeficiency in northern Thailand (1991 to
2011)
Wongkulab P, Wipasa J, Chaiwarith R, Supparatpinyo K (2013), PLOS ONE 8(9): e76371.
 Epidemiology of AOID
2004-2011 are increases
 Most found in Asia.
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6. • Adult-onset immunodeficiency syndrome: AOID (Mortality rate 25%)
• Autoantibody to Interferon gamma (auto-anti IFN-γ)
• Require professional staff
• Labor intensive workflow and time-consuming
• Development of anti-IFN-γ strip test
Enzyme linked immunosorbent assay
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7. Advantage of point of Lateral-flow assay
 Acceptable sensitivity & specificity
 Low cost / test ฿฿฿฿ ฿฿฿฿฿
 Reduced turn around time (min.)
 Early diagnosis
 Improved patient morbidity and mortality rate
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8. 8
Lateral-flow Production Process
Recombinant protein
production
Detection overall of anti-
IFN ϒ by indirect ELISA
Indirect ELISA-Based
IC-strip test for detecting anti-
IFN ϒ in AOID
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interferon-gamma (IFN-γ) monomeric streptavidin (mSV)
Lateral-flow assay
Demonte D., et al. (2014)
Appl Microbiol Biotechnol

9. Titer by indirect ELISA 0 500 2500 12,500
Indirect Strip test - +2 +3 +5
Lateral- Flow assay to Detect Auto anti- IFN-γ in AOID Sera
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10. Sensitivity 73.52%
Specificity 100%
PPV 100%
NPV 78.04%
Sensitivity 84.00%
Specificity 90.24%
PPV 84%
NPV 90.24 %
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Lateral-flow Assay Efficacy Comparison at Different ELISA Cut-off Titer
Significant titer for future clinical sample??????

11. Advantage:
 Novel IC strip platform using in-house materials
 Solving the problem of unsuccessful colloidal gold labeling of IFN-γ
 Benefit for AOID diagnosis
 Suitable for remote hospital
 Improve therapeutic process
Future aspect:
 Evaluate clinical significant with wider collaborative units
 Correlation of antibody titer with disease progression
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Conclusion

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Fabrication of biosensor for Anti-
interferon-gamma antibody detection

13. Fabrication of biosensor for Anti-interferon-gamma antibody detection
Cyclic Voltammetry measurement
by Electrochemical Instruments
 Directly immobilized IFN-γ
 NTA-NI2+-6His tagged protein
sample
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14. Cyclic Voltammetry measurement
by Electrochemical Instruments
Fabricating of biosensor for screening Anti-interferon-gamma antibody in AOID
 Directly immobilized IFN-γ
 NTA-NI2+-6His tagged protein
Serum sample
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15. - Center of biomolecular therapy and diagnostics; CBTD), Faculty of Associated Medical Sciences,
Chiang Mai University (Prof.Dr. Chatchai Tayapiwatana and colleague)
- Center of Innovative Immunodiagnostic Development, Faculty of Associated Medical Sciences,
Chiang Mai University (Prof. Dr. Watchara Kasinrerk and colleague )
- Research Institute for Health Sciences, Chiang Mai University
(Dr. Supachai Sakkhachornphop and colleague)
- Bio-innovation building (Mubio), Mahidol University
Collaborative Research
- i+MED Laboratories Company Limited
Core Research Units
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16. Acknowledgement
 Biomedical Technology Research Center (BMTRC), Faculty of Associated Medical
Sciences, Chiang Mai University
 Research and Researchers for Industry (RRI) Program under the Thailand Research Fund
 Flagship Research Grant from National Research Council of Thailand
 National Science and Technology Development Agency, Thailand; National Research and
Innovation Council, Thailand
 Center of Excellence on Medical Biotechnology: CEMB
 i+MED Laboratories Company Limited, Thailand
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