Novel Field-Based Protein Detection Method Using Light Transmission Spectroscopy and Antibody Functionalized Gold Nanoparticles.

Protein detection is a universal tool critical to many applications in medicine, agriculture, and biotechnology. We developed a novel protein detection method combining light transmission spectroscopy and particle-size analysis of gold nanospheres monovalently functionalized with polyclonal antibodies and applied it to an emerging challenge for such technologies─the monitoring of environmental proteins (eProteins) present in natural aquatic systems. These are an underreported source of pollution and include the pseudopersistent Cry toxins that enter aquatic ecosystems from surrounding genetically engineered crops. The assay is capable of detecting proteins in complex matrices, such as water samples collected in the field, making it a competitive assay for eProtein detection. It is sensitive, reaching 1.25 ng mL-1, and we demonstrate its application to the detection of Cry1Ab from subsurface tile-drain and streamwater samples from agricultural waterways. The assay can also be quickly adapted for other protein detection applications in the future.

Rapid quantitative protein detection by light transmission spectroscopy.

Rapid, sensitive, and quantitative protein detection is critical for many applications in medicine, environmental monitoring, and the food industry. Advancements in detection of proteins include the use of antigen-antibody binding; however, many current methods are time-consuming and have limiting factors such as low sensitivity and the inability to provide absolute values. We present a new high-throughput method for protein detection using light transmission spectroscopy (LTS), which can quantify and size nanoparticles in fluid suspension. LTS can quantify proteins directly and target specific proteins through antigen-antibody binding. This work shows that LTS can distinguish between and quantify bovine serum albumin, its antibody, and the BSA + Ab complex and determine BSA protein concentrations down to 5 µg/mL. We use both Mie and discrete dipole approximation models to provide geometric insight into the binding process.

High-permeability functionalized silicone magnetic microspheres with low autofluorescence for biomedical applications.

Functionalized magnetic microspheres are widely used for cell separations, isolation of proteins and other biomolecules, in vitro diagnostics, tissue engineering, and microscale force spectroscopy. We present here the synthesis and characterization of a silicone magnetic microsphere which can be produced in diameters ranging from 0.5 to 50 µm via emulsion polymerization of a silicone ferrofluid precursor. This bottom-up approach to synthesis ensures a uniform magnetic concentration across all sizes, leading to significant advances in magnetic force generation. We demonstrate that in a size range of 5-20 µm, these spheres supply a full order of magnitude greater magnetic force than leading commercial products. In addition, the unique silicone matrix exhibits autofluorescence two orders of magnitude lower than polystyrene microspheres. Finally, we demonstrate the ability to chemically functionalize our silicone microspheres using a standard EDC reaction, and show that our folate-functionalized silicone microspheres specifically bind to targeted HeLa and Jurkat cells. These spheres show tremendous potential for replacing magnetic polystyrene spheres in applications which require either large magnetic forces or minimal autofluorescence, since they represent order-of-magnitude improvements in each. In addition, the unique silicone matrix and proven biocompatibility suggest that they may be useful for encapsulation and targeted delivery of lipophilic pharmaceuticals.