Abstract
Major advancements in molecular biology and clinical diagnostics cannot be brought about strictly through the use of genomics based methods. Improved methods for protein detection and proteomic screening are an absolute necessity to complement to wealth of information offered by novel, high-throughput sequencing technologies. Only then will it be possible to advance insights into clinical processes and to characterize the importance of specific protein biomarkers for disease detection or the realization of “personalized medicine”. Currently however, large-scale proteomic information is still not as easily obtained as its genomic counterpart, mainly because traditional antibody-based technologies struggle to meet the stringent sensitivity and throughput requirements that are required whereas mass-spectrometry based methods might be burdened by significant costs involved. However, recent years have seen the development of new biodetection strategies linking nucleic acids with existing antibody technology or replacing antibodies with oligonucleotide recognition elements altogether. These advancements have unlocked many new strategies to lower detection limits and dramatically increase throughput of protein detection assays. In this review, an overview of these new strategies will be given.
Highlights
In biological systems, information is passed down from the deoxyribonucleic acid (DNA) to the protein level with RNA as an intermediate
In the field of biosensor research recent years have seen an ever-increasing trend towards the use of deoxyribonucleic acid (DNA)- and ribonucleic acid (RNA) oligonucleotides as mediators in the detection and quantification of protein biomarkers, effectively resulting in a reversal of the flow of information between nucleic acids and proteins
It is clear that the use of DNA labeling can serve to significantly enhance the sensitivity of immunoassays for the detection of protein biomarkers
Summary
Information is passed down from the deoxyribonucleic acid (DNA) to the protein level with RNA as an intermediate. In the field of biosensor research recent years have seen an ever-increasing trend towards the use of deoxyribonucleic acid (DNA)- and ribonucleic acid (RNA) oligonucleotides as mediators in the detection and quantification of protein biomarkers, effectively resulting in a reversal of the flow of information between nucleic acids and proteins The causes for this reversal can be traced back to the realization that major advancements in molecular biology and clinical diagnostics cannot be brought about strictly through the use of modern, generation whole genome sequencing technologies. The fact that ELISA is usually dependent on fluorogenic or chromogenic substrates for actual signal generation will severely limit the number of simultaneous detection events that can be achieved, since the number of distinct dyes as well as the number of detection channels in typical readout instruments is usually limited In this respect, nucleic acid mediated protein assays, where a specific deoxyribonucleic acid (DNA) sequence is coupled to or even wholly constitutes the affinity reagent, might offer some unique benefits [12]:. These will include examples where DNA is used purely for signal amplification and multiplexing in conjunction with conventional antibody recognition elements as well as methods where the olignucleotides themselves (i.e., aptamers) are the recognition elements (Figure 2)
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