Protein Microarray Analysis
Features of Protein Microarray Analysis
Our protein microarray was developed through the Fukushima Translational Research Project. It can comprehensively detect the binding properties of antibodies against over 10,000 different proteins.
Array type
Four types are available: Human proteins (2 types: wheat germ cell-free protein expression system [ENDEXT™ technology], cellular expression system), microorganisms, and allergens. The details of each array can be found below.
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Features of Human Protein Microarray
For human protein microarrays, we provide 2 types of arrays: wheat germ cell-free protein expression system and cellular expression system. A large number of proteins are arrayed and are suitable for profiling circulating antibodies, searching for biomarkers and novel drug targets, profiling of antibody specificity, etc.
Human 18K protein microarray (wheat germ cell-free protein expression system)
Contains more than 16,600 human proteins (representing approximately 70% of all human genes) synthesized using a wheat germ cell-free protein expression system based on ENDEXTTM Technology (CellFree Sciences Co., Ltd.). We offer a wide variety of proteins, ranging from intracellular proteins to secreted proteins and membrane proteins (Fig. A).
The list of the proteins can be downloaded from the following link:
Protein list for human-protein-microarray-wheat-germ-cell-free-protein-system (Excel: 1440KB)
Human 5K protein microarray (cellular expression system)
Contains more than 5,300 human proteins synthesized using cellular expression systems (Fig. B). Compared to the wheat germ cell-free protein expression system, the types of proteins arrayed are limited but suitable for evaluation of post-translational modification and proteins that retain a three-dimensional structure.
The list of the proteins can be downloaded from the following link:
Protein list for human-protein-microarray-expression-system-using-mammalian-cells-etc (Excel: 340KB)
Contents of the proteins by molecular function
Percentage of protein expression systems
Example
Binding specificity analysis can be performed using Human protein microarrays
Specificity and cross reactivity were mapped using commercially available 78 antibodies
Features of Microbial Protein Microarrays
Contains about 1,600 recombinant proteins (normal phase: Fig. E) and 3,400 crude cell lysates (reverse phase: Fig. F) including virus, bacteria, fungi, and protozoa.
The list of the proteins and lysates can be downloaded from the following link:
Protein list for microorganism-protein-microarray_012822 (Excel: 129KB)
Thus, it allows evaluation of the binding property against as many as 5,000 microbial antigens at a time.
Microbial proteins include major viruses such as various coronaviruses, influenza viruses, and adenoviruses as well as dengue viruses, Zika viruses, and Ebola viruses, pathogenic bacteria such as Escherichia coli and Staphylococcus aureus, and tropical malaria plasmodium and toxoplasma as protozoa.
Example
1,606 recombinant proteins in total are spotted on a microarray
3,432 lysates in total are spotted on a microarray
Before vaccination, few individuals had antibodies to the SARS-CoV-2 proteins, but after vaccination, antibodies binding to SARS-CoV-2 antigens were uniformly found. On the other hand, there was almost no change in the profile of antibodies to SARS coronavirus, also indicating that this microarray is highly reproducible.
The antibodies against reversed-phase lysates (33 spots) of influenza viruses were shown as a heat map. Antibodies against various strains were identified in sera from healthy individuals. Also, the cross-reactivity of commercial antibodies to the virus strains are evaluated.
Features of Allergen Protein Microarray
Includes reverse phase samples (crude) of potential allergens ranging from foods such as grains, fruits, and seafood, and materials such as cotton and latex to plants, pollen, mites, and bacteria. It allows for assessment of antibody responses to as many as 1,200 potential allergen candidates at once (Fig. I) with a very small amount of sample.
1,195 crude allergens in total are spotted on a microarray
Example
Reactive spots are extracted from the results of IgE profiling using plasma from 5 allergic patients and 4 healthy subjects. The heat map makes it easy to identify IgEs to specific allergens.
Features of Bioinformatics analysis
For efficient and high-quality data analysis, we developed a bioinformatics analysis system, TUNiNG-BASE™.
TUNiNG-BASE™ equips various functions like below;
- Multiple normalization methods
- PCA analysis for data quality test
- Clustering analysis
- Machine learning
- Pathway/Disease enrichment analysis
Example
A) PCA analysis: TUNiNG-BASE™ can normalize data using 5 different methods. Example of PCA analysis revealing data bias between batches B) Various parameters can be entered and a list of those that meet the criteria, as well as a box-and-whisker diagram can be shown C) List of interest can be viewed in terms of diseases and pathways through Enrichment analysis D) The list can also be subjected to clustering analysis to see associations between samples or between antigen proteins
Data deliverables
- Analysis data (Excel format)
- Test report (PDF format)
Sample requirements
The types of samples and antibodies capable of measuring differ between organism species. In addition, the volume of solution required for measurement depends on the type of sample. Volumes are shown below. Please contact us for more details.
| Serum, Plasma, Milk (mammal), Saliva | > 100 μL |
|---|---|
| Cerebrospinal fluid, Urine, Semen | > 300 μL |
| Lacrimal fluid | > 50 μL |
| Antibody solution (> 1 mg/mL) | > 20 μL |
Price
Please contact us first.