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Conformational Evaluation of HIV-1 Trimeric Envelope Glycoproteins Using a Cell-based ELISA Assay
Published on: September 14, 2014
Using microcantilever deflection to detect HIV-1 envelope glycoprotein gp120
Yee Lam1, Nehal I Abu-Lail, Munir S Alam
1Department of Mechanical Engineering and Materials Science, Duke University, Durham, North Carolina 27708, USA.
Nanomedicine : Nanotechnology, Biology, and Medicine
|February 13, 2007
Summary
Microcantilever sensors detect HIV-1 gp120 by measuring deflection caused by antibody binding. This shows potential for a portable, low-cost HIV-1 detection device.
Area of Science:
- Biotechnology
- Nanotechnology
- Immunology
Background:
- Microcantilevers are sensitive biosensors that detect biomolecules through deflection.
- Specific binding events on microcantilever surfaces induce measurable stress and bending.
Purpose of the Study:
- To utilize microcantilever deflection for detecting human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) gp120.
- To demonstrate the sensor's capability in detecting specific antibody-gp120 interactions and induced-fit binding.
Main Methods:
- Decorating microcantilevers with monoclonal antibodies (mAbs) A32 or T8 to capture gp120 from solution.
- Measuring cantilever deflection upon gp120 binding and subsequent incubation with mAb 17b.
Main Results:
- Observed significant cantilever deflection upon specific binding of gp120 to A32- or T8-coated cantilevers.
- Demonstrated further deflection increase with mAb 17b on A32-presenting cantilevers, indicating induced-fit interaction detection.
- Achieved detection at test concentrations of 8 microg/mL gp120 and 0.17 mg/mL 17b.
Conclusions:
- Microcantilever deflection sensors can effectively detect HIV-1 gp120 and specific binding interactions.
- The technique shows promise for identifying induced-fit binding events relevant to HIV-1.
- Further development could yield a portable, cost-effective diagnostic tool for HIV-1 detection.
