Using microcantilever deflection to detect HIV-1 envelope glycoprotein gp120.
Microcantilevers have been used over the last decade to detect biomolecules from solution. Specific binding events on one surface of the microcantilever create a differential stress, resulting in measurable deflection. Here we use this principle to detect human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) gp120 from solution. We observed deflections approximately twice that of the baseline (in PBS) upon specific binding of gp120 to cantilevers decorated on one side with monoclonal antibodies (mAbs) A32 or T8. Subsequent incubation with mAb 17b (known to bind an A32-induced epitope on gp120) further increased deflection of A32- but not T8-presenting cantilevers. This work shows the capability of microcantilever deflection sensors to detect an induced-fit interaction at test concentrations of 8 microg/mL gp120 and 0.17 mg/mL 17b. Further development of this technique could lead to a portable, low-cost device for the effective detection of HIV-1.
Lam
Yee
Y
Department of Mechanical Engineering and Materials Science, Duke University, Durham, North Carolina 27708, USA.
Abu-Lail
Nehal I
NI
Alam
Munir S
MS
Zauscher
Stefan
S
D000911:Antibodies, Monoclonal / Q000737:chemistry; D000939:Epitopes; D015483:HIV Antibodies / Q000737:chemistry; D015699:HIV Envelope Protein gp120 / Q000737:chemistry*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D011092:Polyethylene Glycols / Q000737:chemistry; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D013997:Time Factors; D014616:Vaccinia virus / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D011092:Polyethylene Glycols