Antibody Profiling
Antibody Profiling - Related Videos
12:19
Antibody Profiling by Luciferase Immunoprecipitation Systems (LIPS)
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Cited by 146 •
2009
The technical aspects of performing LIPS (Luciferase Immunoprecipitation Systems) are described. The overall approach involves expressing chimeric genes encoding antigens fused to Renilla luciferase (Ruc) in mammalian cells. Crude Ruc-antigen extracts are then prepared and, without purification, employed in immunoprecipitation assays to quantify antibodies.
05:21
A Peptide Microarray Technology for Specificity Profiling of Antibodies
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2025
This video demonstrates a technique that applies peptide microarray technology to profile the specificity of antibodies recognizing post-translational modifications (PTMs) on histone peptides. A microarray slide containing immobilized fluorophore-conjugated histone peptides with known PTM combinations is treated with target PTM-specific antibodies. This process helps identify the antibody's specificity for the target and its potential cross-reactivity with non-target PTMs.
15:17
Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing
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Cited by 19 •
2011
We described a procedure for the disaggregation of colorectal cancer (CRC) to produce viable single cells, which are then captured on customized antibody microarrays recognizing surface antigens (DotScan CRC microarray). Sub-populations of cells bound to the microarray can be profiled by fluorescence multiplexing using monoclonal antibodies tagged with fluorescent dyes.
13:21
Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
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Cited by 6 •
2012
In this study, we describe an improved protocol for a multiplexed high-throughput antibody microarray with lectin detection method that can be used in glycosylation profiling of specific proteins. This protocol features new reliable reagents and significantly reduces the time, cost, and lab equipment requirements as compared to the previous procedure.
11:08
Chemo-enzymatic Synthesis of N-glycans for Array Development and HIV Antibody Profiling
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Cited by 2 •
2018
A modular approach to the synthesis of N-glycans for attachment to an aluminum oxide-coated glass slide (ACG slide) as a glycan microarray has been developed and its use for the profiling of an HIV broadly neutralizing antibody has been demonstrated.
FAQs
It examines several antibody characteristics rather than relying on abundance alone. Specificity indicates which antigens antibodies recognize, affinity describes the strength of those interactions, and isotype identifies the antibody class. Considering these features together can reveal whether a sample contains a broad, focused, strong, or class-specific pattern of immune recognition.
The interaction used in the assay determines which antibody population becomes measurable. Selected antigens can test recognition of particular targets, whereas antigen-specific reagents can capture antibodies with defined binding properties. Consequently, the choice of binding strategy shapes the immune-recognition pattern observed and determines which aspects of the sample are emphasized.
These approaches measure antibody-related information through different experimental formats. Protein arrays examine interactions across selected protein targets, immunoassays quantify binding using assay-based detection, and sequencing-based approaches investigate antibody repertoire information through sequence data. The method therefore influences whether the resulting profile emphasizes antigen recognition, measured binding, or repertoire-related patterns.
Abundance indicates how much of an antibody-related signal is present, but it does not describe the full response. Affinity adds information about interaction strength, while isotype identifies the antibody class involved. Including these measurements helps distinguish samples that may have similar quantities yet differ in binding behavior or antibody composition.
A typical workflow begins with a biological sample and a defined set of antigen targets or antigen-specific capture reagents. Antibody interactions are then measured using a protein array, immunoassay, or sequencing-based approach. The resulting data are characterized according to features such as abundance, specificity, affinity, and isotype to identify immune-response patterns.
Method selection should follow the biological question and the antibody features that need to be characterized. Protein arrays and immunoassays support measurements based on antigen binding, whereas sequencing-based approaches provide a route to examine antibody repertoire changes. Matching the assay format to the desired readout helps produce information relevant to the study objective.
Researchers apply it to compare immune responses associated with infections, vaccines, tumors, and autoimmune conditions. The resulting profiles can support biomarker discovery, evaluation of vaccine efficacy, and investigation of how antibody repertoires change during disease or treatment. These applications connect molecular antibody measurements with broader changes in biological or clinical states.