Epigenetics is the study of heritable changes in gene expression that do not involve alterations to the DNA sequence. These changes can be influenced by various factors, including DNA methylation, histone modifications, and non-coding RNAs (especially microRNAs). These epigenetic marks encompass the epigenome, which is natural and critical to many organism functions, although significant negative effects can happen if epigenetic processes occur improperly.1,2
Histone post-translational modifications (PTMs) include acetylation, methylation, phosphorylation, and ubiquitylation on histone tails. Histone proteins are important in packaging DNA, so their PTMs can greatly affect the activation or repression of gene transcription. Since histone residues can have none or several different modifications, identifying them can be challenging.3
Epigenetic antibodies can be used in chromatin immunoprecipitation (ChIP) experiments to detect and characterize epigenetic targets. They can be either polyclonal or monoclonal antibodies and can be produced with high specificity to a particular PTM and ensure consistent antibody lot-to-lot variability. This is valuable as identifying the correct genomic position and quantity of histone PTMs will allow the study of their downstream functions.4
Besides histone modifications, DNA methylation is a well-studied epigenetic process, characterized by the addition of a methyl or hydroxymethyl group to the C5 position of the cytosine. DNA methylation plays a role in regulating gene expression and several biological activities including aging and disease.5 Antibodies targeting this can be used in techniques such as dot blot, enzyme-linked immunosorbent assay (ELISA), methylated DNA immunoprecipitation (MeDIP), immunofluorescence (IF), and immunohistochemistry (IHC).
Non-coding RNAs are a cluster of RNAs that do not encode functional proteins, but ongoing research supports them as being important in epigenetic control, regulating gene expression. Non-coding RNAs include small interfering RNAs (siRNAs), microRNAs (miRNAs), PIWI-interacting RNAs (piRNAs), and long non-coding RNAs (lncRNAs). These RNAs appear to interact in a regulatory network, but analysis their relationships remain a challenge.6 To analyze RNA-protein interactions, researchers can use techniques involving antibodies such as RNA Immunoprecipitation (RIP) and UV Cross-Linking and Immunoprecipitation (CLIP).
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Weinhold, B. (2006). Epigenetics: The Science of Change. Environmental Health Perspectives, 114(3), A160. https://doi.org/10.1289/ehp.114-a160
Li, G., Zan, H., Xu, Z., & Casali, P. (2013). Epigenetics of the antibody response. Trends in Immunology, 34(9), 460. https://doi.org/10.1016/j.it.2013.03.006
Mariño-Ramírez, L., Kann, M. G., Shoemaker, B. A., & Landsman, D. (2005). Histone structure and nucleosome stability. Expert Review of Proteomics, 2(5), 719. https://doi.org/10.1586/14789450.2.5.719
Nishikori, S., Hattori, T., Fuchs, S. M., Yasui, N., Wojcik, J., Koide, A., Strahl, B. D., & Koide, S. (2012). Broad Ranges of Affinity and Specificity of Anti-Histone Antibodies Revealed by a Quantitative Peptide Immunoprecipitation Assay. Journal of Molecular Biology, 424(5), 391-399. https://doi.org/10.1016/j.jmb.2012.09.022
Moore, L. D., Le, T., & Fan, G. (2013). DNA Methylation and Its Basic Function. Neuropsychopharmacology, 38(1), 23-38. https://doi.org/10.1038/npp.2012.112
Wei, J., Huang, K., Yang, C., & Kang, C. (2017). Non-coding RNAs as regulators in epigenetics (Review). Oncology Reports, 37, 3-9.https://doi.org/10.3892/or.2016.5236
The therapeutic efficacy of antibodies is closely related to their ability to recognize and bind specific epitopes on target antigens. Epitopes, or antigenic determinants, are a group of amino acids or other chemical groups that are part of a molecule to which an antibody attaches itself. Epitope characterization can help reveal the mechanism of antibody binding and apply intellectual property (patent) protection for novel antibodies, in addition to designing antibodies with high specificity and minimal cross-reactivity.
Understanding the differences between antibody specificity and selectivity is essential for designing and interpreting antibody-based assays in research for experimental accuracy and data interpretation. Antibody specificity refers to an antibody's ability to recognize and bind to a particular epitope—a unique part of an antigen that elicits an immune response.
Antibody-based assays are essential tools in biomedical research, providing the means to detect, quantify, and visualize specific proteins or antigens within complex biological samples. These assays' efficacy hinges on the antibodies' precise properties. While affinity, avidity, specificity, and selectivity are fundamental to antibody performance, the ultimate impact of these properties is heavily influenced by the experimental context in which the antibody is employed.
Biologics, particularly antibodies, have become indispensable in biomedical research and therapeutic development. Research-use-only (RUO) biologics play a pivotal role in preclinical studies, providing researchers with the necessary tools to explore antibody functions and therapeutic potential in vivo.
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