Multiplex Analysis of Autoantibodies
Detecting autoantibodies is critical in various research areas, such as cytokine research, infectious disease research, autoimmune disease research, and cancer research. Multiplex analysis of autoantibodies, such as MILLIPLEX® multiplex assays, helps advance this research by detecting multiple analytes at once to give researchers a full autoantibody profile.
Section Overview
- What Are Autoantibodies?
- Multiplexing Autoantibodies
- Dual Reporter Autoantibody Profiling
- Measuring Cytokine Autoantibodies
- Autoimmune Autoantibody Analysis
- Cancer Autoantibody Detection
- Neurodegenerative Autoantibody Detection
- Related Products
- Select Publications Using MILLIPLEX® Human Autoantibody Multiplex Kits
What Are Autoantibodies?
Autoantibodies are antibodies that react with substances/antigens that are self-produced. This happens when the body cannot tell the difference between its own antigens and invading antigens. These autoantibodies lead to inflammation and the destruction of tissues and organs. They are often associated with autoimmune disorders but can be found in other diseases as well.
Examples of autoantibodies include:
- Cytokine autoantibodies
- Autoimmune autoantibodies
- Cancer autoantibodies
- Neurodegenerative disease autoantibodies
Multiplexing Autoantibodies
Multiplex analysis of autoantibodies helps researchers save time in the lab by measuring multiple autoantibodies in one sample. With high-quality multiplexing like MILLIPLEX® multiplex panels using the Luminex® platform, researchers have the option of analyzing a variety of autoantibodies as well as the flexibility to choose which analytes they want to assay. Figure 1 shows the capture antigen method these assays use.
MILLIPLEX® autoantibody kits are for qualitative measurement of autoantibodies in serum and plasma samples. Each kit contains four assay controls beads to be mixed with the selected antigen-immobilized beads and a PE-Ig conjugate as the detection component.
Figure 1.The MILLIPLEX® autoantibody panel assay depiction. This represents a capture antigen method.
Dual Reporter Autoantibody Profiling
Profiling both IgG and IgM autoantibodies in biological samples provides valuable insights into the immune response and potential disease conditions. The MILLIPLEX® Dual Reporter Autoantibody IgG/IgM Reagent Pack offers an innovative solution for serological research by simultaneously detecting IgG and IgM autoantibodies in a single assay. This exclusive Dual Reporter Pack leverages advanced two-laser technology of the xMAP® INTELLIFLEX DR-SE System to measure two antibody types on a single bead, providing insights into both recent (IgM) and chronic (IgG) immune responses. This reagent pack serves as a companion product to our expanding range of MILLIPLEX® serology assays or use it with your own antigen-immobilized beads.
Figure 2.The MILLIPLEX® Dual Reporter Reagent Pack enables detection of both IgG and IgM antibodies.
Measuring Cytokine Autoantibodies
Measuring cytokine autoantibodies may be useful in disease research since anti-cytokine antibodies occur frequently and are present in healthy and patient samples with various infections, acquired immunodeficiency, or autoimmune diseases. Cytokines offering protection against microbes can be targeted by cytokine autoantibodies, leading to life-threatening infections. Measuring cytokine autoantibodies may be useful for research on disease monitoring and the efficacy of treatment.
Cytokine Autoantibody Multiplex Analysis
With multiplex analysis of cytokine autoantibodies, you can measure more biomarkers in one sample. Our MILLIPLEX® Human Cytokine Autoantibody Expanded IgG Panel contains antigen-coated beads for the simultaneous detection of 23 autoantibodies in serum or plasma samples. Four assay control beads are included as part of the base kit, and you can select the cytokine autoantibodies you want to assay as a part of your experiment. Figure 2 shows example data of using this panel in autoimmune disease and sepsis samples, compared to healthy samples.
Figure 3.Using Product Number HCYTABG-17K, the MFI (Median Fluorescence Intensity) values of the cytokine autoantibody biomarkers were determined in autoimmune disease, including systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) (n=19), sepsis (n=17), and healthy (n=9) serum samples (diluted 1:100). Data from 4 select analytes in the kit are shown.
Autoimmune Autoantibody Analysis
Autoantibody formation is principal to the pathogenesis of a variety of autoimmune diseases. Dysregulated apoptosis and the subsequent defective clearance of cellular debris lead to the exposure of autoantigens and the generation of autoantibodies. The presence of autoantibodies may indicate disease activity, prognosis, and clinical associations related to a variety of autoimmune diseases, including systemic lupus erythematosus (SLE), Sjögren’s Syndrome, Systemic Sclerosis, Polymyositis (PM)/Dermatomyositis, and various overlap syndromes of these diseases. Table 1 describes autoimmune autoantibodies and their disease associations.
Autoimmune Autoantibody Multiplex Analysis
The MILLIPLEX® Human Autoimmune Autoantibody Panel contains 20 antigens for detection in serum or plasma samples. These autoantibodies include:
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Figure 4 shows a comparison of disease and normal samples using this panel.
Figure 4.Using Product Number HAIAB-10K, the Median Fluorescence Intensity (MFI) was determined for 19 disease samples – 10 systemic lupus erythematosus, 7 granulomatosis with polyangiitis, and 2 polymyositis, and 10 normal samples. Disease samples were much higher for 14 of the 20 analytes in this panel.
Figure 5 shows autoantibody MFI values compared to clinical laboratory data, showing that this panel followed similar trends to what was measured by a commercial lab.
Figure 5.Clinical laboratory data were available for 6 autoantibodies (SSA/Ro60, SSA/Ro52, SSB/La, RNP, Sm, and Scl-70) in 8 SLE patients. Those SLE patients positive by clinical laboratory methods also demonstrated elevated MFI in the Autoimmune Autoantibody panel. The SLE patients considered negative by the same clinical laboratory methods exhibited lower MFI.
Figure 6 describes the reference serum analysis with this panel.
Figure 6.Reference sera were available for evaluation of 10 autoantibodies in the MILLIPLEX® Human Autoimmune Autoantibody Panel. The controls exhibited elevated MFI for the autoantibodies for which they were expected to react strongly.
This shows the value of using multiplex technology to evaluate multiple autoantibodies, allowing for an extensive autoantibody profile to evaluate samples from various autoimmune diseases.
Cancer Autoantibody Detection
Cancer autoantibodies to tumor-associated antigens are being studied as potential cancer biomarkers, as well as possible reporters of early carcinogenesis and indicators of cancer prognosis. Research is being done to find accurate, reliable, and affordable blood tests that are needed for the early detection of cancer.
Cancer Autoantibody Multiplex Analysis
The MILLIPLEX® Human Cancer Autoantibody Panel contains cancer antigen-coated beads for simultaneous detection of 15 autoantibodies in serum or plasma samples. It offers the ability to select the cancer autoantibodies needed for your experiment. Figure 7 shows a comparison between cancer and normal samples using this panel.
Figure 7.Using Product Number HCABMAG-13K, the MFI (Median Fluorescence Intensity) values of the cancer autoantibody biomarkers were determined in cancer (breast, colorectal, ovarian, lung, prostate) (n=78) and healthy control (n=18) serum samples (samples diluted at 1:100).
Figure 8 describes the reactivity of serial diluted serum cancer autoantibody samples.
Figure 8.The cancer autoantibody multiplex assay is robust and requires less than 5 µL of human serum samples, as the sera were tested at 1:100 dilution. Note twelve of the fifteen analytes are shown.
This human cancer autoantibody multiplex assay can be used alone or in combination with other MILLIPLEX® cancer circulating biomarker panels, as useful biomarker screening tools in studying the early detection of cancer.
Neurodegenerative Autoantibody Detection
Neurodegenerative autoantibodies target critical components of the nervous system including synaptic proteins, neuronal antigens, and cell surface receptors. This binding disrupts normal neural signaling and elicits both inflammatory responses and tissue damage, driving the progression of various neurological disorders. Using these autoantibodies as biomarkers can provide critical insight into neuroinflammation and identify potential therapeutic options in preclinical research.
Neurodegenerative Autoantibody Multiplex Analysis
The MILLIPLEX® Human Neurodegenerative Autoantibody Panel delivers high-throughput screening by simultaneously detecting up to 25 IgG autoantibodies in multiple sample types including serum, plasma, and cerebrospinal fluid (CSF). Pairing with the MILLIPLEX® Dual Reporter Autoantibody IgG/IgM Reagent Pack and the xMAP® INTELLIFLEX DR-SE System doubles the datapoints per sample by detecting both IgG and IgM autoantibodies in a single workflow. Figure 9 illustrates differences between the IgG and IgM autoantibody MFI values for selected analytes in presumed healthy controls and samples from individuals with neurodegenerative conditions.
Figure 9.IgG and IgM autoantibody MFIs in 16 presumed healthy controls and 40 neurodegenerative disease [stroke, multiple sclerosis (MS), Alzheimer’s disease (AD), and Parkinson’s disease (PD)] serum and plasma samples diluted 1:100. Significant differences were determined by Mann-Whitney test (*P < 0.05, **P < 0.01) although this may not indicate disease.
For Research Use Only. Not For Use In Diagnostic Procedures.
Select Publications Using MILLIPLEX® Human Autoantibody Multiplex Kits
Product No.: HCYTABG-17K
- COVID-19 associated autoimmunity is a feature of severe respiratory disease - a Bayesian analysis
- Toll-like Receptor Homologue CD180 Ligation of B Cells Upregulates Type I IFN Signature in Diffuse Cutaneous Systemic Sclerosis
Product No.: HAIAB-10K
- Distinct Autoimmune Antibody Signatures Between Hospitalized Acute COVID-19 Patients, SARS-CoV-2 Convalescent Individuals, and Unexposed Pre-Pandemic Controls
- Pathological consequences of anti-citrullinated protein antibodies in tear fluid and therapeutic potential of pooled human immune globulin-eye drops in dry eye disease
- IRF5 genetic risk variants drive myeloid-specific IRF5 hyperactivation and presymptomatic SLE
- In vivo efficacy of combined human broadly neutralizing antibodies against hepatitis B virus
- Distinct circulating autoantibodies are associated with COVID-19 hospitalization and SARS-CoV-2 neutralization activity
- Autoantibodies in COVID-19 convalescent plasma donors
- Longitudinal analysis reveals elevation then sustained higher expression of autoantibodies for six months after SARS-CoV-2 infection
Product No.: HCABMAG-13K
- Prognostic Value of Baseline Autoantibodies in Metastatic NSCLC Patients Receiving PD-/PDL-1 Targeted Immunotherapy
- A Luminex Approach to Develop an Anti-Tumor-Associated Antigen Autoantibody Panel for the Detection of Prostate Cancer in Racially/Ethnically Diverse Populations
For Research Use Only. Not For Use In Diagnostic Procedures.
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