|Schematic from a single well of NanoInk's 48 sub-array 10 plex cytokine assay. HTS format can detect multiple biomarkers simultaneously from as little as 2 μL of sample
|Illustration of nanoscale protein analysis assay
|Protein nanoarray data vs. conventional protein microarray data
Dip Pen Nanolithography® (DPN®) is ideal for printing proteins at micron and sub-micron scales. DPN deposition does not subject biological materials to destructive forces (such as compression, shear, harsh vacuum, or UV light) that can damage structure and function, so it is a highly biocompatible process. High viscosity liquids (from 1 to 20,000 cP) are also simple to print using the DPN platform. NanoInk systems are especially adept at simultaneously depositing multiple biomolecules (including antibodies) in nanoscale patterns, delivering the ability to miniaturize and multiplex protein analysis.
Disadvantages of Conventional Protein Arrays
Traditional pin-spotting protein microarray technologies often print inconsistent features, leading to poor assay reproducibility. Ink jet printing is known to aerosolize reagents during the array printing process, which can cause splattering, higher localized backgrounds and lower signal to noise ratios. Traditional multi-well ELISAs and bead-based protein analysis techniques typically require large amounts of sample material and reagents and can exhibit prohibitively slow reaction kinetics.
Benefits of DPN
DPN patterns exhibit highly uniform and repeatable features, leading to exceptional assay reproducibility. Unlike conventional ELISA and bead-based assays, nanoarray assays require just 2 µl of sample so are able to generate multiplexed proteomic biomarker data from rare and precious samples like spinal fluid, rodent serum, tumor extracts, tears, and dried blood spots. DPN also delivers rapid reaction kinetics within small reagent volumes, substantially lowering assay costs compared to traditional immunoassays. NanoInk’s miniaturized assay format, coupled with the 0.5 µm resolution detection capabilities of the NanoArray Assay System scanner, results in up to single femtogram/ml assay sensitivity. As a result, NanoInk’s nanoscale immunoassay platform can detect and quantitate even low abundance protein biomarkers from a wide variety of sample types.
NanoInk protein analysis assays require just 2μl of sample
The Nano BioDiscovery Division’s protein array-based instrument systems, assay kits, and contract services are uniquely well-suited for high-throughput drug discovery, diagnostic development, and proteomic research applications like:
- Biomarker detection and quantification
- Cytokine expression
- Toxicity screening
- Dried blood spot analysis
- Angiogenic factor screening
- Growth factor and signal transducer screening
- Apoptosis protein screening
- Protease screening
- Chemokine and adipokine screening
- Antibody titer screening
Multiplexed Protein Arrays
- Development and Validation of Ultra High Sensitive and Low Volume Multiplex Rat Renal Toxicity Protein NanoArray Assay
Describes the development and proof of concept testing of a nanoarray assay capable of detecting albumin, beta-2 microglobulin, clusterin, cystatin C, KIM-1, and trefoil factor 3 in the single digit pg/ml range.
- Miniaturized Multiplex Highly Sensitive Protein NanoArray Assay for Measuring Biomarkers in Dried Blood Spot (DBS) Samples
Presents development and validation data, as well a proprietary elution protocol, for a DBS-sample based assay that can generate maximum biomarker data with minimum sample volumes.
- Multiplexed Highly Sensitive Protein NanoArray Assays for Small Volume Samples
Details development of a nanoscale sandwich ELISA platform that exhibits improved sensitivity to detect both cytokine and angiogenesis biomarkers in the 100's of fg/ml range from as little as 2-4 µl of sample.
- Miniaturized Multiplex Highly Sensitive Protein NanoArray Assay for Small Volume Serum and Tumor Samples (2-4 µl)
Explains validation of a nanoscale assay for the sensitive detection and accurate quantification of low-abundance protein biomarkers in human and mouse serum samples, human and mouse tumor samples, and DBS samples.
- High Sensitivity Biomarker Detection Using Protein NanoArrays
Presents benefits of detecting biomarkers using a new nanoarray-based platform capable of generating nano to micron-scale features of biomolecules on any solid surface.
- High Sensitive Cytokine Detection Multiplex Assay Using Nanoarrays
Describes optimization of DPN technology to deposit nanoarrays of human inflammation cytokine antibodies that retain high sensitivity toward their respective antigens and then conduct multiplex protein analysis on these arrays.
- Immobilization and Detection of Proteins on the Nanoscale
Provides data on the development of nanoscale antibody and kinase assays using DPN technology.
- Protein NanoArrays: Extending the Limits of Biomarker Detection
Details proof-of-concept study on a nanoscale kinase assay for three growth factor receptors: Her2 receptor, epidermal growth factor receptor, and insulin-like growth factor-1 receptor.
- Fabrication of Protein and Oligomer Nanoarrays
Presents data and challenges associated with the nanoscale printing of high-density DNA and protein arrays using a DPN technique.
- High Sensitivity and Small Sample Volume Protein Detection Assays Using Multiplex Nanoarrays
Describes development of a 10-plex human inflammation and a 10-plex human angiogenesis ELISA using the ultrasensitive nanoscale array platform.
Protein Immobilization and Biomarker Detection
Details use of the DPN platform to generate highly uniform and repeatable micron to nanoscale sized features of multiple proteins on a variety of substrates.
1. J.-W. Jang; A. Smetana; P. Stiles. Multiplexed Dip Pen Nanolithography Patterning by Simple Desktop Nanolithography Platform. Scanning, 31 (2010) 1-6.