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Abstract
Detection of nucleic acids using antibodies is uncommon. This is in part because nucleic acids are poor immunogens and it is difficult to elicit antibodies having high affinity to each type of nucleic acid while lacking cross-reactivity to others. We describe the origins and applications of a variety of anti-nucleic acid antibodies, including ones reacting with modified nucleosides and nucleotides, single-stranded DNA, double-stranded DNA, RNA, DNA:RNA hybrids, locked-nucleic acids or peptide nucleic acid:nucleic acid hybrids. Carefully selected antibodies can be excellent reagents for detecting bacteria, viruses, small RNAs, microRNAs, R-loops, cancer cells, stem cells, apoptotic cells and so on. The detection may be sensitive, simple, rapid, specific, reproducible, quantitative and cost-effective. Current microarray and diagnostic methods that depend on cDNA or cRNA can be replaced by using antibody detection of nucleic acids. Therefore, development should be encouraged to explore new utilities and create a robust arsenal of new anti-nucleic acid antibodies.
Acknowledgements
The authors thank Susan Gottesman and Gisela Storz of the NIH (Bethesda, MD, USA) for critical discussions on this topic during our work together and crucial comments during preparing for this article. Z Hu is in debt to Jacques Ravel of the University of Maryland (Baltimore, MD, USA) for providing software and Affymetrix genomic microarray chips of Bacillus anthracis for detection of non-coding RNAs, and to Noel Gerald (Silver Spring, MD, USA) and Stephanie Healey (Winchester, MA, US) of the US FDA for useful suggestions. Due to limited space we apologize to our colleagues that their invaluable works could not be included in this publication.
Disclaimer
Some antibodies and other research tools described here are available from commercial sources. We are not recommending a particular product, but only providing an example of where it might be available. The authors do not stand for official FDA and NIH opinions of the biologic, drug or device products mentioned in this article.
Financial & competing interests disclosure
Research described here was supported in part by Oak Ridge Institute for Science and Education, USA, and by the Intramural Research Program of the NIH. The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.
No writing assistance was utilized in the production of this manuscript.
Key issues
Immunoassays for detection of nucleic acids as antigens are not frequently used. This is most likely because:
– PCR and sequencing methods have high sensitivity and specificity.
– Nucleic acids are poor immunogens, making it difficult to elicit antibodies specific to different types of nucleic acids.
– The anti-nucleic acid antibodies created decades ago are now largely neglected.
Anti-nucleic acid antibodies include types that react with modified nucleosides and nucleotides, single-stranded DNA, double-stranded DNA, single-stranded RNA, double-stranded RNA, DNA:RNA hybrids, locked-nucleic acids or peptide nucleic acid:nucleic acid hybrids and others.
Anti-hybrid antibodies, combined with hybrid capture and PCR-EIA, are used for simple and rapid detection of bacteria and viruses.
– The most notable applications are the US FDA-approved Qiagen’s Digene Hybrid Capture technologies for in vitro diagnostic tests of Neisseria gonorrhoeae, Chlamydia trachomatis and HPV.
Current microarray systems have problems detecting low concentrated small non-coding RNAs due to the multiple steps involved in labeling and synthesis of cDNA or cRNA.
– Antibody-based microarray detection is sensitive and unbiased since synthesis of cDNA or cRNA is avoided. It was successfully applied for bacterial sRNA detection, yeast transcriptome mapping and cancer detections from FFPE samples.
Anti-nucleic acids antibodies, combined with biosensors, ChIP/DRIP, sequencing and immunofluorescence, are used for successful detections of miRNAs, R-loops, metakaryotic stem cells and apoptotic cells.
Anti-nucleic acid antibodies, together with modern technologies, can lead to the expanding or upgrading of current methods widely used in clinical labs and biological research.
