DNA aptamer affinity ligands for highly selective purification of human plasma-related proteins from multiple sources

Abstract

Nucleic acid aptamers are promising ligands for analytical and preparative-scale affinity chromatography applications. However, a full industrial exploitation requires that aptamer-grafted chromatography media provide a number of high technical standards that remained largely untested. Ideally, they should exhibit relatively high binding capacity associated to a very high degree of specificity. In addition, they must be highly resistant to harsh cleaning/sanitization conditions, as well as to prolonged and repeated exposure to biological environment. Here, we present practical examples of aptamer affinity chromatography for the purification of three human therapeutic proteins from various sources: Factor VII, Factor H and Factor IX. In a single chromatographic step, three DNA aptamer ligands enabled the efficient purification of their target protein, with an unprecedented degree of selectivity (from 0.5% to 98% of purity in one step). Furthermore, these aptamers demonstrated a high stability under harsh sanitization conditions (100h soaking in 1M NaOH). These results pave the way toward a wider adoption of aptamer-based affinity ligands in the industrial-scale purification of not only plasma-derived proteins but also of any other protein in general.

Application of aptamers for in vivo molecular imaging and theranostics

“…Aptamers have provided fundamental insights concerning the versatility of RNA, supporting its involvement in the origin of life [14,15]. In parallel, aptamers have been increasingly used for biotechnological applications, such as biosensors [16], bio-purification [17], biomarker discovery [18], and the regulation of gene expression [19]. Furthermore, one aptamer has already been commercialized as a drug for the treatment of age-related macular degeneration and several others are currently in clinical trials [20].…”

Aptamers as the Agent in Decontamination Assays (Apta-Decontamination Assays): From the Environment to the Potential Application In Vivo

“…Apart from biosensing application, aptamers have also been the subject in the aptamer-based capture assay. In the capture assay applications, aptamers are applied for the purification of their corresponding targets for a wide variety of purposes [ 11 , 27 , 28 ]. The present review seeks to present an overview of the more narrowed context of the aptamer-based capture assay, which is on the usage of the aptamer in the decontaminating process or simply abbreviated as apta-decontamination assay.…”

Programmable and Multifunctional DNA‐Based Materials for Biomedical Applications

“…Given high binding capacity in combination with very high degree of specificity, DNA aptamers are promising ligands for not only analytical but preparative‐scale affinity chromatography applications. Forier et al exemplified aptamer affinity chromatography for the purification of three human therapeutic proteins whose purity increases from 0.5% to 98% within one step . Comparing to the traditional Ni‐NTA‐based protein purification technology, aptamer affinity chromatography resulted in a 3.6‐fold higher l ‐selectin yield with increased purity …”