Autoantibody Microarray
The autoantibody microarray can detect a wide variety of self-antigens, such as cell nuclear components, cytoplasmic proteins, phospholipid proteins, cell matrix proteins, mucosal/secreted proteins, glomeruli, and other tissue-specific proteins. It has been widely used for screening of autoantigen specificities associated with different manifestations of diseases. Equipped with advanced technologies and experienced scientists, Creative Biolabs can provide a whole set of natural autoantibodies (NAA) detection and analysis services for our worldwide customers, especially with the help of autoantibody microarray platform. Our state-of-art technology platform and rich expertise for NAA services will be helpful for your disease diagnosis, treatment or drug design and development projects.
Background of Autoantibody
Autoantibodies are a type of antibodies produced by the immune system that are directed against one or more of the individual's own proteins also called autoantigens. Autoantibodies are associated with many autoimmune diseases (notably lupus erythematosus). Normally, autoantibodies can be eliminated by the immune system’s self-regulatory process routinely via the neutralization of autoantibody-producing lymphocytes before they mature. If this process fails, antibodies will react to self-components resulting in diseases occurrence. Autoantibodies elicit body tissues damage through the phagocytosis (ingestion) or lysis (bursting) of healthy cells. Especially the blood cells become the main targets of these actions. For instance, autoimmune hemolytic anemia is caused by autoantibodies binding to red blood cells. In addition, autoantibodies harbor the ability to interfere with the normal cells. For example, in Graves’ disease, autoantibodies bind to receptor cells of the thyroid gland, which stimulates the overproduction of thyroid hormones.
Fig.1 Mechanisms for autoantibody production: apoptosis, antigen modification, and cross-reactivity. (Suurmond, 2015)
Autoantibody Microarray
Autoantibodies are associated with various autoimmune diseases and the dysregulation of the immune system. Profiling of autoantibodies against a wide range of autoantigens is a powerful tool for the identification of biomarkers and for prediction and diagnosis of diseases. Currently, the autoantibody microarray can be used to detect the IgG, IgM, IgA and IgE autoantibodies from serum or other body fluids.
- Autoantibody Microarray Service for IgM
IgM autoantibody from serum or other body fluids can be detected by using the autoantibody microarray. The samples will be treated with DNAse I, and then incubated with autoantigen array. The autoantibodies binding to the antigens on the array will be detected with labeled anti-IgM and the arrays will be scanned with microarray scanner. The images will be analyzed using software to generate GPR files. The averaged fluorescent intensity (NFI) of each autoantigen will be normalized to internal controls. The final data production will be an Excel spreadsheet containing Non-normalized and Normalized signal intensity as well as the Signal to Noise Ratio (SNR) for all antigens.
- Autoantibody Microarray Service for IgG
IgG autoantibody has 4 subclasses (IgG1, IgG2, IgG3 and IgG4) and each of them may be associated with different clinical manifestations during the disease development. With a high-throughput autoantigen microarray platform, we are able to detect subclasses of IgG autoantibodies against a broad range of autoantigens with high sensitivity and specificity. With this assay, we can simultaneously detect 4 IgG subclasses against various autoantigens on each array. The samples will be treated with DNAse before incubated with autoantigen array. The autoantibodies binding to the antigens on the array will be assayed with biotinylated and IgG secondary antibody and then detected with fluorescent-labeled anti-biotin for imaging. The arrays will be scanned with Microarray Scanner and images will be analyzed using special software to generate GPR files. The averaged fluorescent intensity (NFI) of each autoantigen will be normalized to internal controls.
- Autoantibody Microarray Service for IgA and IgE
IgA and IgE are also important autoantibody subtypes for many autoimmune, infection and allergic diseases. With our high-throughput autoantigen microarray platform, a variety of autoreactive IgA and IgE autoantibodies can be detected with high sensitivity and specificity. In this service, we can simultaneously detect autoreactive IgA and IgE autoantibodies against up to over 100 antigens on each array. The samples will be treated with DNAse I, then incubated with autoantigen array. The autoantibodies binding to the antigens on the array will be detected with biotinylated and IgA and IgE secondary antibody and then detected with fluorescent-labeled anti-biotin for imaging. The arrays will be scanned with Microarray Scanner and images will be analyzed using special software to generate GPR files. The averaged fluorescent intensity (NFI) of each autoantigen will be normalized to internal controls (IgA or IgE).
Fig.2 A detailed representation of ‘reverse capture’ autoantibody microarray. Based on the platform of ELISA, this method uses an antibody microarray to capture native antigens. Autoantibody reactivity is then evaluated by differentially labeling patient IgG and incubating these antibodies with the native antigens that are immobilized on the antibody microarray. (Ehrlich, 2006)
Features
- High throughput, and multiplicity of the whole workflow
- High sensitivity, high specificity, and high accuracy to detect NAA
- Competitive and affordable price
As a professional service provider in antibody analysis, design and preparation, Creative Biolabs offers comprehensive NAA services to help you detect NAA associated immune diseases and new NAA biomarkers using autoantibody microarray in a timely and cost-effective manner. Our tailored services and high-quality products will contribute greatly to the success of your projects. Please contact us for more information and a detailed quote.
References:
- Suurmond, J.; Diamond, B. Autoantibodies in systemic autoimmune diseases: specificity and pathogenicity. Journal of Clinical Investigation. 2015, 125(6): 2194.
- Ehrlich, J.R.; et al. The 'reverse capture' autoantibody microarray: a native antigen-based platform for autoantibody profiling. Nature Protocols. 2006, 1(1): 452-60.