Antibody Cross-Reactivity Analysis
Understanding antibody cross-reactivity
Cross-reactivity refers to the ability of an antibody to bind to different antigens that share similar epitopes. This phenomenon occurs when the epitopes on different antigens have enough structural similarity for an antibody to recognize and bind to both.
Why Cross-reactivity matters
- Research context: In experimental research, cross-reactivity can lead to false-positive results or misinterpretation of data, impacting the reliability of experimental outcomes.
- Therapeutic context: For therapeutic antibodies, cross-reactivity can cause unintended side effects by binding to off-target proteins, potentially leading to decreased therapeutic efficiency or adverse reactions.
- Diagnostic context: In diagnostic applications, cross-reactivity can compromise the specificity of assays, resulting in inaccurate diagnostics.
Profiling cross-reactivity against human proteins
PEPperPRINT offers multiple solutions for analyzing antibody specificity and cross-reactivities against human proteins, human B-cell epitopes, or even other disease- or pathogen-specific antigens.
- Protein-level screening: Straigtforward detection and analysis of protein targets with the HuProtTM Human Proteome Microarray(currently only available for customers based in Europe).
- Human epitome-wide screening: Identification of reactivity against known human B-cell epitopes with the PEPperCHIP® Autoimmune Human Epitope Microarray or PEPperCHIP® Human Epitome Microarray.
- Broad epitope signature screening: Detection of cross-reactivites against other disease- or pathogen-specific antigens or random peptides with PEPperCHIP® Custom Peptide Microarrays.
Cross-reactivity screening of COVID-19 patient sera against known human autoantigens. Study design (left) and representative microarray scans (right) of the PEPperCHIP® Autoimmune Human Epitope Microarray showing different cross-reactivity profiles across different patient cohorts.
| HuProtTM Proteome Microarray | PEPperCHIP® Human Epitome Microarray | PEPperCHIP® Autoimmune Human Epitope Microarray | |
|---|---|---|---|
| Microarray content | over 21,000 human proteins | 29,128 peptides covering all linear B-cell epitopes of the Immune Epitope Database | 3,723 human B-cell epitopes associated with autoimmune diseases |
| Species | Human | Human, animal, and various pathogens | Human |
| Peptide length | - | 5 to 17 amino acids depending on epitope length; longer epitopes translated into overlapping peptides | 15 amino acids |
| Peptide overlap shift | - | shift +1 | shift +1 |
| Sample material | 10-50 µL serum or 5-30 µg purified antibody | 100-200 µl serum or 100-200 µg purified antibody | 10-20 µl serum or 10-20 µg purified antibody |
| Analysis | Available by PEPperMAP® Service only | Available by PEPperMAP® Service only | DIY following the PEPperCHIP® Immunoassay Protocol, or by PEPperMAP® Service |
Antibody cross-reactivity use cases
Antibody cross-reactivity analysis is a crucial step in establishing antibody specificity and determining general antigenic targets. Learn how antibody cross-reactivity analysis with PEPperCHIP® Peptide Microarrays can accelerate your research by exploring some of our selected case studies and customer publications.
Application notes
- Antibody cross-reactivity profiling of a human monoclonal antibody with the PEPperCHIP® Human Epitome Microarray
- Antibody validation with a combination of protein and peptide microarrays
Poster
Cited publications
- Analysis of Plasmablasts From Children With Kawasaki Disease Reveals Evidence of a Convergent Antibody Response to a Specific Protein Epitope (Rowley et al. 2023)
- Differential patterns of cross-reactive antibody response against SARS-CoV-2 spike protein detected for chronically ill and healthy COVID-19 naïve individuals (Jaago et al. 2022)
- A cross-reactive mouse monoclonal antibody against rhinovirus mediates phagocytosis in vitro (Behzadi et al. 2020)