Protein Array for Gut Microbial Biomarker Identification

The human gut microbiome is composed of numerous microbes performing functions that affect human health. Based on a cutting-edge Gut Microbiota-Based Diagnostics Platform, Creative Biolabs offers biomarker identification services using protein array technology. Our knowledge and expertise allow us to assist with the discovery of gut microbial biomarkers while supporting decision-making and mitigating risk at key stages of your program.

Protein Arrays

Protein arrays use protein probes that in solution retain the ability to interact specifically with other proteins or molecules. Protein arrays can be used to identify protein-protein interactions, enzyme-substrate interactions, and antibody-antigen interactions. Based on their fabrications and applications, protein arrays can be classified into three major categories: analytical, functional, and reverse-phase protein arrays. Protein array allows simultaneously monitoring hundreds of thousands of protein samples in cells or tissues. At present, the analytical protein array has been applied to identify biomarkers for a wide variety of diseases such as cancers.

Fig.1 Classification of three types of protein arrays. (Huang, 2021)

Our Services

Protein array has enabled large-scale screening of proteins in different biosamples that are applicable for biomarker discovery and allow for the simultaneous and rapid analysis of thousands of proteins in high-throughput. These arrays have been produced by expressing and purifying proteins, which are then printed on the array surface. However, many challenges accompany this approach including the difficulty in purifying many proteins. Thus, we take an alternate approach, to construct by spotting protein-encoding plasmid DNA at high density, in an addressable fashion, on an array surface. Each array is printed with recombinant proteins. This approach avoids problems with protein purification and stability. Then, the nascent protein arrays are used for protein binding analysis. The workflow is as follows:

• Plasmid DNA is mixed with psoralen-biotin and cross-linked using ultraviolet light.
• The DNA is mixed with avidin/streptavidin, crosslinker, and the anti-glutathione S-transferase (GST) capture antibody, and this mix is arrayed on the slides.
• After blocking, a cell-free expression mix is applied to the slide, and during a temperature-programmed incubation, the proteins are produced and bind to the capture antibody.
• The slide is washed, and the proteins are detected by detecting the GST tag.

Creative Biolabs is participating in microbial biomarker identification by applying our infrastructure for sample collection, data processing, and integrated analysis. We have many strategies to identify microbial biomarkers including direct mass spectrometry, immunoprecipitation & mass spectrometry, and protein array. If you have any questions about our service, please contact us for more information.

References

1. Huang, Y.; Zhu, H. Protein array-based approaches for biomarker discovery in cancer. Genomics, proteomics & bioinformatics. 2017, 15(2): 73-81.
2. Ramachandran, N.; et al. On-chip protein synthesis for making microarrays. New and Emerging Proteomic Techniques. Humana Press. 2006, 328: 1-14.

For Research Use Only | Not For Clinical Use