Gold nanoparticles have garnered interest as both radiosensitzers and computed tomography (CT) contrast agents. However, the extremely high concentrations of gold required to generate CT contrast is far beyond that needed for meaningful radiosensitization, which limits their use as combined therapeutic–diagnostic (theranostic) agents. To establish a theranostic nanoplatform with well-aligned radiotherapeutic and diagnostic properties for better integration into standard radiation therapy practice, a gold- and superparamagnetic iron oxide nanoparticle (SPION)-loaded micelle (GSM) is developed. Intravenous injection of GSMs into tumor-bearing mice led to selective tumoral accumulation, enabling magnetic resonance (MR) imaging of tumor margins. Subsequent irradiation leads to a 90-day survival of 71% in GSM-treated mice, compared with 25% for irradiation-only mice. Furthermore, measurements of the GSM-enhanced MR contrast are highly predictive of tumor response. Therefore, GSMs may not only guide and enhance the efficacy of radiation therapy, but may allow patients to be managed more effectively.
mRNA display is a powerful method for in vitro directed evolution of
polypeptides, but its time-consuming, technically demanding nature has hindered its widespread use.
We present a streamlined protocol in which lengthy mRNA purification steps are replaced with faster
precipitation and ultrafiltration alternatives; additionally, other purification steps are entirely
eliminated by using a reconstituted translation system and by performing reverse transcription after
selection, which also protects input polypeptides from thermal denaturation. We tested this
procedure by performing affinity selection against Her2 using binary libraries containing a
non-specific designed ankyrin repeat protein (DARPin) doped with a Her2-binding DARPin (dopant
fraction ranging from 1:10 to 1:10,000). The Her2-binding DARPin was recovered in all cases, with an
enrichment factor of up to two orders of magnitude per selection round. The time required for one
round is reduced from 4–7 days to 2 days with our protocol, thus simplifying and
accelerating mRNA display experiments.
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