Protein Biochip Technology Platform (Multi Marker)
Near-Infrared Fluorescent Labeled Chip
Silica-Based Membrane as Protein Chip
What is Protein Biochip Technology?
Protein chip is a new type of biochip, which is composed of protein microarrays fixed on different types of support media. The position and composition of the fixed molecules in the array are known. Unlabeled or labeled biological molecules react with the probes on the chip, and then detect them through a specific scanning device, and the results are analyzed and processed by a computer. Protein chips have the following characteristics:
- High specificity
- High sensitivity
- High throughput
- Good repeatability
- Strong applicability
How Do We Customize Protein Biochips?
Protein chips can be divided into functional research chips and detection chips according to their functions. Functional research chips are mostly high-density chips, with natural proteins or fusion proteins fixed on the carrier. This type of chip is mainly used for protein activity and proteomics related research; the density of detection chips is relatively low, and antigens, antibodies, etc. are fixed, which are mainly used for large-scale and rapid detection of biological molecules.
Figure 1. Schematic representation of the new AD multiplex protein biochip. (Sources: Veitinger M, et al. 2014)
- Choice of Carrier
The carrier of protein chips is basically the same as that of gene chips, mainly including: filter membrane, gel and glass slide. Among them, filter membrane and gel have the advantages of large protein fixation amount, high protein activity and the ability to provide three-dimensional space for protein fixation, but these carriers often cannot meet the requirements of high mechanical spotting strength of proteins, and the samples spotted on them are prone to diffusion, resulting in mutual interference between different samples. Glass slides have been widely used in the production of protein chips due to their smooth surface, low cost and stable performance However, in this type of fixation method, the proteins fixed on the glass slide are in different directions, so some proteins cannot react biologically with the corresponding components in the sample to be tested in the subsequent process due to the lack of free reaction domains.
- Preparation of Probe Protein
Protein chips require a large number of probes with high affinity and strong specificity to be fixed on the carrier. Probes can be selected from different antigens, antibodies, enzymes and receptors according to different research purposes. Due to its high specificity and affinity, monoclonal antibodies are a better probe protein. Chips constructed with them can be used to detect the expression abundance of proteins and identify new proteins.
- Protein Chip Detection
Currently, there are two different detection methods for the detection of protein chip reaction results: the first is direct detection, that is, directly detecting the captured target protein. This type of method includes surface enhanced nanocluster resonance technology, surface plasmon resonance (SPR) for detecting changes in refractive index, solid-phase laser excitation time-resolved fluorescence spectroscopy, surface enhanced laser desorption ionization-time of flight mass spectrometry (SELDI-TOF-MS), etc.
Table 1. Comparison of protein chip technology and traditional protein interaction detection technology.
| Feature/Aspect | Protein Biochip Technology | Traditional Protein-Protein Interaction Detection Technologies |
| Detection Principle | Uses microarrays to analyze multiple protein interactions simultaneously | Methods include co-immunoprecipitation, yeast two-hybrid, affinity chromatography, pull-down assays, FRET, SPR, BRET, cross-linking, far-Western blotting, and protein microarrays |
| Sample Throughput | High throughput; capable of analyzing thousands of interactions at once | Lower throughput; usually analyzes one interaction per experiment |
| Sensitivity | High sensitivity due to amplification and signal detection mechanisms | Variable sensitivity; depends on the specific method used |
| Specificity | High specificity with well-optimized arrays and conditions | Variable specificity; often requires extensive optimization |
| Quantification | Provides quantitative data on protein interactions and expression levels | Often semi-quantitative; quantitative data can be obtained with additional steps |
| Miniaturization | Highly miniaturized; requires small sample volumes | Less miniaturized; generally requires larger sample volumes |
| Multiplexing Capability | High; can simultaneously detect and analyze multiple proteins | Limited; typically one or a few proteins at a time |
| Automation and Speed | Highly automatable and fast; can process many samples in parallel | Often manual and time-consuming; each interaction requires separate setup |
| Data Analysis | Advanced bioinformatics tools for large-scale data analysis | Less complex data; easier but more labor-intensive analysis |
| Cost | Initial setup can be expensive, but cost-effective for large-scale studies | Generally lower initial cost, but can become expensive for large-scale studies |
| Applications | High-throughput screening, biomarker discovery, diagnostics, drug development | Basic research, validation of interactions, functional studies |
| Dynamic Range | Wide dynamic range for detecting protein abundance and interaction strength | Variable dynamic range depending on the method |
| Reproducibility | High reproducibility with standardized protocols and conditions | Variable reproducibility; can be influenced by experimental conditions |
| Technological Complexity | Requires specialized equipment and expertise | Generally less complex; widely available equipment and protocols |
Application of Protein Chips
- Proteomics research
- Enzyme-substrate interaction research
- Anti-HIV research
- Screening and detection of tumor markers
References
- Zhu H, Snyder M. Protein chip technology. Curr Opin Chem Biol. 2003, 7(1):55-63.
- Cretich M, et al. Protein microarray technology: how far off is routine diagnostics? Analyst. 2014, 139(3):528-42.
- Dupuy AM, et al. Protein biochip systems for the clinical laboratory. Clin Chem Lab Med. 2005, 43(12):1291-302.
- Huang W, et al. New Insights into the Tumor Microenvironment Utilizing Protein Array Technology. Int J Mol Sci. 2018 Feb 13;19(2):559.
- Veitinger M, et al. A platelet protein biochip rapidly detects an Alzheimer's disease-specific phenotype. Acta Neuropathol. 2014, 128(5):665-77.