Abstract
Polyethylene glycol (PEG) conjugation (PEGylation) is a well-established strategy to improve the pharmacokinetic and immunogenic properties of a wide variety of nanomedicines and therapeutic peptides and proteins. This broad use makes PEG an attractive ‘allround’ candidate marker for the biodistribution of such PEGylated compounds. Here, we present the development of a novel strategy for PEG quantification in biological matrices. It is based on sample hydrolysis which both decomposes the sample matrix and degrades PEGylated analytes to specific molecular fragments more suitable for detection by LC-MS/MS. Method versatility was demonstrated by applying it to a wide variety of PEGylated compounds, including polymeric poly(ethylbutyl cyanoacrylate) (PEBCA) nanoparticles, lipidic nanoparticles (Doxil®, LipImage 815™ and lipid nanoparticles for nucleic acid delivery) and the antibody Cimzia®.
Method applicability was assessed by analyzing plasma and tissue samples from a comprehensive drug biodistribution study in rats, of both PEBCA containing the anticancer drug cabazitaxel, and LipImage 815™ nanoparticles containing the near-infrared dye IR780-oleyl. The results demonstrated the method’s utility for biodistribution studies on PEG. Importantly, by using the method described herein in tandem with quantification of nanoparticle payloads, we showed that this approach can provide detailed understanding of various critical aspects of the in vivo behavior of PEGylated nanomedicines, such as drug release and particle stability. As seen from Figure 1, there is a very notable difference between how the polymeric and lipidic nanoparticles behave in liver tissue, when comparing the measured concentrations of nanocarrier base material (quantified as PEG) versus the encapsulated – and subsequently released – payload.
Together, the presented results demonstrate the novel method as a robust, versatile and generic approach for biodistribution analysis of PEGylated therapeutics.
Method applicability was assessed by analyzing plasma and tissue samples from a comprehensive drug biodistribution study in rats, of both PEBCA containing the anticancer drug cabazitaxel, and LipImage 815™ nanoparticles containing the near-infrared dye IR780-oleyl. The results demonstrated the method’s utility for biodistribution studies on PEG. Importantly, by using the method described herein in tandem with quantification of nanoparticle payloads, we showed that this approach can provide detailed understanding of various critical aspects of the in vivo behavior of PEGylated nanomedicines, such as drug release and particle stability. As seen from Figure 1, there is a very notable difference between how the polymeric and lipidic nanoparticles behave in liver tissue, when comparing the measured concentrations of nanocarrier base material (quantified as PEG) versus the encapsulated – and subsequently released – payload.
Together, the presented results demonstrate the novel method as a robust, versatile and generic approach for biodistribution analysis of PEGylated therapeutics.
