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Aptamers as a sensitive tool to detect subtle modifications in therapeutic proteins
Ran Zichel1, Wanida Chearwae, Gouri Shankar Pandey
1Laboratory of Hemostasis, Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.
Plos One
|March 3, 2012
Summary
DNA aptamers can detect subtle conformational changes in therapeutic proteins, even when activity is unaffected. This novel approach offers a routine method for drug development and quality control, providing early warnings of structural alterations.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Protein Science
Background:
- Therapeutic proteins can undergo subtle conformational changes during production, impacting safety and efficacy.
- Existing bioanalytical methods struggle to detect these tertiary structure alterations.
- Current techniques like X-ray crystallography and NMR are impractical for routine quality control.
Purpose of the Study:
- To demonstrate DNA aptamers as surrogate antibodies for detecting protein conformational changes.
- To establish aptamers as a viable tool for drug development and quality control.
- To provide a novel analytical method for monitoring protein structural integrity.
Main Methods:
- Developed and utilized DNA aptamers targeting thrombin.
- Employed ELISA and label-free platforms for aptamer-based assays.
- Replicated heat-treatment to induce conformational changes in thrombin.
Main Results:
- A panel of DNA aptamers successfully detected conformational changes in thrombin.
- Aptamers identified alterations even when protein specific activity remained unaffected.
- The aptamer-based assay proved more sensitive than traditional antibody methods in this context.
Conclusions:
- DNA aptamers offer a sensitive and practical approach to monitor protein conformational changes.
- This method can be integrated into routine drug development and quality control.
- Aptamers provide an early warning system for manufacturing-induced structural variations, preventing adverse outcomes.
