A carbon nanotube-based Raman-imaging immunoassay for evaluating tumor targeting ligands.
Herein, we describe a versatile immunoassay that uses biotinylated single-walled carbon nanotubes (SWNTs) as a Raman label, avidin-biotin chemistry to link targeting ligands to the label, and confocal Raman microscopy to image whole cells. Using a breast tumor cell model, we demonstrate the usefulness of the method to assess membrane receptor/ligand systems by evaluating a monoclonal antibody, Her-66, known to target the Her2 receptors that are overexpressed on these cells. We present two-dimensional Raman images of the cellular distribution of the SWNT labels corresponding to the distribution of the Her2 receptors in different focal planes through the cell with validation of the method using immunofluorescence microscopy, demonstrating that the Her-66-SWNT complexes were targeted to Her2 cell receptors.
Department of Chemistry, The University of Texas at Dallas, Richardson, TX 75080-3021, USA. imusselm@utdallas.edu.
Pooja
P
Bajaj
Carole
C
Mikoryak
Ruhung
R
Wang
David K
DK
Bushdiecker
Pauras
P
Memon
Rockford K
RK
Draper
Gregg R
GR
Dieckmann
Paul
P
Pantano
Inga H
IH
Musselman
D045744:Cell Line, Tumor; D005455:Fluorescent Antibody Technique; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008024:Ligands; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D037742:Nanotubes, Carbon; D009369:Neoplasms / Q000378:metabolism*; D013057:Spectrum Analysis / Q000379:methods*
D008024:Ligands; D037742:Nanotubes, Carbon