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Image of DPN-generated nanoarray composed of 4 different antibodies, each tagged with a
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
Illustration of nanoscale protein analysis assay
Illustration of nanoscale protein analysis assay
Protein nanoarray data vs. conventional protein microarray data

Protein Analysis

Overview

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
NanoInk protein analysis assays require just 2μl of sample

Applications

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/discovery
  • Cytokine expression profiling
  • Angiogenic factor screening
  • Toxicity screening
  • Growth factor and signal transducer screening
  • Apoptosis protein screening
  • Protease screening
  • Chemokine and adipokine screening

Application Notes

Multiplexed Protein Arrays

Posters

Miniaturized Multiplex Highly Sensitive Protein NanoArray Assay for Measuring Biomarkers in Dried Blood Spot (DBS) Samples
Development and Validation of Ultra High Sensitive and Low Volume Multiplex Rat Renal Toxicity Protein NanoArray Assay
Multiplexed Highly Sensitive Protein NanoArray Assays for Small Volume Samples
Miniaturized Multiplex Highly Sensitive Protein NanoArray Assay for Small Volume Serum and Tumor Samples (2-4 µl)

High Sensitivity Biomarker Detection Using Protein NanoArrays
High Sensitive Cytokine Detection Multiplex Assay Using Nanoarrays
Immobilization and Detection of Proteins on the Nanoscale
Protein NanoArrays: Extending the Limits of Biomarker Detection
Fabrication of Protein and Oligomer Nanoarrays
High Sensitivity and Small Sample Volume Protein Detection Assays Using Multiplex Nanoarrays
Protein Immobilization and Biomarker Detection

Publications/References

1.    J.-W. Jang; A. Smetana; P. Stiles. Multiplexed Dip Pen Nanolithography Patterning by Simple Desktop Nanolithography Platform. Scanning, 31 (2010) 1-6.

 

 

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