Protein microarray (protein arrays or protein biochips) technology is a process of immobilizing numerous proteins on to a solid support and using miniaturized assays to describe protein interactions, that has been developed steadily in recent years. It is a powerful methodology, in a high-throughput format, to analyze thousands of known proteins in a single experiment. Protein microarrays are now an indispensable tool for proteomic applications, multi-parameter clinical diagnostic tests, and many other fields.

Protein Microarray

Applications in Basic Research:

  • Development of new assays
  • Detection of protein-binding properties (Protein-protein interaction, protein-peptide interaction, protein-DNA interaction, protein-small molecule interaction, protein-glycan interaction)
  • Profiling monoclonal antibody specificity
  • Protein posttranslational modifications (PTMs) (Protein phosphorylation, protein ubiquitylation, protein acetylation, S-nitrosylation)

Applications in Clinical Research:

  • Biomarker identification
  • Host-pathogen interactions

As a well-recognized partner in the drug development process, Creative Biolabs has launched a series of versatile microarray platform and offers three major types of protein microarrays to study protein function and construction of networks. The arrays containing customized sets of proteins, as well as array profiling services, are also available for our worldwide clients.

  • How Does Array Work?

Microarray technology refers to the miniaturization of a large number of assays on one small plate. Protein microarray has been prevalently used to capture and measure proteins from biologic samples in a high-throughput style. Typically, a protein microarray consists of a small piece of glass or plastic coated with thousands of capture reagents matching specific proteins. This technology allows it possible to isolate and study many potential biomarker proteins for drug discovery. Protein microarrays can be miniaturized and arranged in a grid with tens of thousands of capture features, each specific for a given protein. Thus, they are considered a multiplexed device and can be tested for multiple markers simultaneously, which is essential for research, diagnostics, and monitoring disease states.

  • Protein Microarray Platforms at Creative Biolabs

Biomarker identification using protein microarrays. Fig.2 Biomarker identification using protein microarrays. (Zhu, 2012)

Protein microarray is an emerging technology that has been viewed to overcome the limitation of DNA microarrays. This tool is excellent for characterizing hundreds of thousands of proteins in a high-parallel and high-throughput fashion. At Creative Biolabs, we provide three major categories of protein microarrays to illustrate their applications in both basic and clinical research. Each array contains superior-quality purified proteins spotted onto the glass slide. Our robust protein microarrays and popular services contain items including but not limited to:

  • Analytical Protein Microarrays - They’re mostly represented by antibody microarrays and can be used to profile a complex mixture of proteins in order to measure binding affinities, specificities, and protein expression levels. In this technique, a library of antibodies or aptamers is arrayed on a glass slide and the targeted protein will be detected either by direct labeling or a reporter antibody in sandwich assay format.
  • Functional Protein Microarrays - They’re constructed using individually purified proteins that involve full-length functional proteins or protein domains. This class of array enables the study of biochemical properties of proteins, such as binding activities (including protein-protein, protein-DNA, protein-peptide, protein-drug, and protein-lipid interactions) and enzyme-substrate relationships through various types of biochemical reactions.
  • Reverse-phase Protein Microarrays (RPAs) - They employ an opposite array format by immobilizing many different lysate samples (tissue, cell lysates, or fractionated cell lysates) on the same chip. And many different probes can be tested to specifically identify specific proteins in these lysate samples. RPAs are frequently detected with fluorescent, chemiluminescent, or colorimetric assays, allowing for the determination of post-translational modifications to define altered proteins as a result of the disease.
  • Fabrication of Protein Microarrays at Creative Biolabs

Fabrication of high-content functional protein microarrays. Fig.3 Fabrication of high-content functional protein microarrays. (Zhu, 2012)

Fabrication of protein microarrays remains many challenges in some critical aspects, particularly in producing proteins in a high-throughput way. At Creative Biolabs, we have established the following services to generate abundant amounts of proteins, which ensure the quality of the resultant protein microarrays.

  • Protein Production
  • Analytical protein microarrays -- Hybridoma-based technology, recombinant antibody technology, phage display, etc.
  • Functional protein microarrays -- Saccharomyces cerevisiae (yeast) protein expression system, Escherichia coli (E. coli) protein expression system, etc.
  • Reverse-phase protein microarrays -- Isolated from cell culture; frozen, ethanol-fixed, or paraffin-embedded tissues; laser captured microdissections of cell populations from certain tissues.
  • Surface Chemistry

Protein immobilization on a solid support is also crucial, following protein production. An ideal surface for array fabrication has to be capable of protein immobilization and preserving 3D conformation of proteins. Here, we provide several common surfaces for microarray fabrication as below.

System Surface
Covalent immobilization Aldehyde, Epoxy, NHS, Carboxylic Ester, etc.
Adsorption Polyvinylidene fluoride (PVDF), Nitrocellulose membrane, Polystyrene
Diffusion Agarose/polyacrylamide Gel, Hydrogel
Affinity capture Ni2+-NTA, Streptavidin, Glutathione
Metal Gold, Silver, Steel, etc.
  • Signal Detection

In label-dependent detections, we have developed several types of labeling reagents, such as fluorescent dyes (Cy3, Cy5, and equivalents), enzymes, liposomes, and radioisotopes.

In label-free detections, no processes affect protein activity and a real-time measurement for the dynamics of protein interactions can be obtained. We’d like to introduce these detection methods, including surface plasmon resonance spectroscopy (SPR), imaging optical ellipsometry (OE), reflectometric interference spectroscopy (RIFS), and oblique-incidence reflectivity difference (OIRD) technique.

  • Advantages
  • Protein microarrays can assay up to hundreds of proteins at a time
  • High-throughput capability for processing up to 16 samples in parallel on each slide
  • The arrayed results can be collected in little as 1 day
  • Small sample volume (as little as 2-5 µL of serum) is needed for detection

Protein microarrays have become a promising approach for a wide variety of applications, especially the identification of small molecule drugs. With rich experience and advanced technologies, Creative Biolabs offers multiple easily customizable protein microarrays to analyze hundreds to thousands of known proteins in proteomics and diagnostic fields. If you want more information, please don’t hesitate to contact us.

Reference

  1. Zhu, H.; et al. Applications of functional protein microarrays in basic and clinical research. Adv Genet. 2012, 79: 123-155.

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