What is Human Milk Oligosaccharide Microarray?
Human milk oligosaccharides (HMOs) are a diverse and structurally complex group of carbohydrates found abundantly in human breast milk. They play crucial roles in infant nutrition, immune modulation, and the establishment of healthy gut microbiota. The human milk oligosaccharide microarray is a high-throughput analytical tool designed to profile and characterize these oligosaccharides systematically. By employing this microarray technology, researchers can elucidate the intricate patterns and functions of HMOs, advancing our understanding of their biological roles.
Principle of Human Milk Oligosaccharide Microarray
An HMO microarray is a high-throughput platform that allows researchers to investigate the interactions between HMOs and different biological samples, such as proteins, antibodies, cells, cell lysates, serum, vesicles, bacteria, or viral particles. The key principle behind an HMO microarray is the immobilization of a diverse set of structurally defined HMOs on a solid surface, typically a glass slide.
Each HMO microarray slide contains multiple identical subarrays, with each subarray featuring a collection of different HMO structures. These HMO structures are covalently attached to the slide surface, creating a high-density array of oligosaccharides.
To study the binding interactions, the biological sample of interest (e.g., a protein or bacterial culture) is labeled with a fluorescent dye or other detectable marker. This labeled sample is then incubated with the HMO microarray, allowing the sample to interact with the immobilized HMOs. After washing away any unbound material, the microarray is scanned to detect the fluorescent signals, which indicate the binding of the sample to specific HMO structures.
Human Milk Oligosaccharide Microarray Platform Offered by Creative Proteomics
High-throughput Screening: The platform enables the simultaneous screening of a large number of HMO samples, allowing researchers to efficiently identify specific structures or patterns of interest.
Comprehensive Analysis: Creative Proteomics offers comprehensive analysis services, including structural characterization, quantification, and functional studies of HMOs.
Customization: The platform can be customized to meet the specific needs of researchers, including the design of microarrays tailored to their research objectives.
Expert Support: Creative Proteomics' team of experts provides guidance and support throughout the entire analysis process, from experimental design to data interpretation.
Advanced Technologies: The platform incorporates advanced analytical technologies and methodologies to ensure accurate and reliable results.
Advantages of Our Human Milk Oligosaccharide Microarray Platform
- Exceptional Sensitivity: Detects oligosaccharides at picomolar levels, enabling precise characterization of complex samples.
- Comprehensive Coverage: Offers a vast library of oligosaccharide probes, ensuring accurate profiling of diverse HMO structures.
- Quantitative Precision: Provides precise measurement of HMO concentrations in milk samples, aiding in understanding variations and impacts on composition.
- Efficient High Throughput: Rapidly screens large sample volumes, ideal for studies requiring swift analysis of diverse samples.
- Flexible Customization: Tailors assays to specific research needs, facilitating targeted experiments.
- Reliable Results: Built on robust technology with stringent quality control, ensuring dependable and reproducible data.
Workflow of Human Milk Oligosaccharide Microarray Assay
Sample Preparation
Human milk samples are collected and pretreated by spiking with an internal standard (e.g., raffinose) to correct for sample losses during processing. HMOs are extracted from the milk samples using solid-phase extraction over C18 and graphite microcolumns.
Fluorescent Labeling
The extracted HMOs are fluorescently labeled with a dye like 2-aminobenzamide (2AB) by incubating at elevated temperature (e.g., 65°C for 2 hours). Unreacted dye is removed by another solid-phase extraction step over silica microcolumns.
Microarray Printing
The HMO microarray contains a comprehensive set of structurally defined HMO structures, typically around 50 different HMOs. Each microarray slide has 16 identical sub-arrays, allowing simultaneous analysis of multiple samples.
Sample Incubation
The labeled HMO samples are incubated on the microarray slide, allowing binding interactions between HMOs and the biological sample of interest (e.g., proteins, antibodies, bacteria).
Detection and Imaging
After incubation, unbound materials are washed away. The microarray is scanned using a fluorescence microarray scanner at the appropriate wavelength for the labeling dye (e.g., 532nm for Cy3). The fluorescence signals indicate the binding of specific HMOs to the biological sample.
Data Analysis
The fluorescence intensity data is analyzed to determine the binding patterns and selectivity of the biological sample towards different HMO structures. This provides insights into the functional roles of HMOs and their interactions with proteins, pathogens, or other biomolecules.
Sample Requirements for Human Milk Oligosaccharide Microarray Assays
| Sample Type | Sample Volume |
|---|---|
| Human Milk | 50-200 μL |
| Serum/Plasma | 100-500 μL |
| Urine | 100-500 μL |
| Saliva | 50-200 μL |
| Breast Tissue | 10-50 mg |
| Cell Culture Media | 500 μL - 1 mL |
| Fecal Samples | 50-200 mg |
| Amniotic Fluid | 200-500 μL |
| Cord Blood | 100-500 μL |
| Synovial Fluid | 200-500 μL |
| Cerebrospinal Fluid | 100-500 μL |
Applications of Human Milk Oligosaccharide Microarray
Breastfeeding Studies: Investigating the composition and variations of HMOs in human milk from different populations and lactation stages.
Infant Health and Development: Understanding the role of HMOs in infant nutrition, immune development, and microbiome modulation.
Gut Microbiota Research: Studying the interactions between HMOs and gut microbiota, including prebiotic effects and modulation of microbial composition.
Disease Biomarker Discovery: Identifying HMO biomarkers associated with diseases such as metabolic disorders, gastrointestinal diseases, and immune-related conditions.
Nutritional Supplements: Screening and selecting specific HMOs for inclusion in infant formula and nutritional supplements to mimic the benefits of human milk.
Biomedical Research: Investigating the molecular mechanisms underlying the biological activities of HMOs, including anti-adhesive, anti-inflammatory, and antimicrobial properties.
Diagnostic Assays: Developing diagnostic assays based on HMO profiles for assessing maternal health, infant immune status, and susceptibility to infections.
Therapeutic Interventions: Exploring the therapeutic potential of HMOs in the prevention and treatment of diseases, including gastrointestinal disorders and infectious diseases.
Personalized Nutrition: Tailoring nutritional interventions based on individual HMO profiles to optimize health outcomes and support personalized medicine approaches.
Environmental and Sociodemographic Factors: Investigating the influence of environmental factors, maternal diet, lifestyle, and sociodemographic factors on HMO composition and health outcomes.