Attachment of poly(ethylene glycol) (PEG) to proteins, peptides, liposomes, drugs, and nanoparticles can improve pharmaceutical pharmacokinetic properties and enhance in vivo biological efficacy. Since the first PEGylated product was approved by the Food and Drug Administration in 1990, increasing numbers of PEGylated compounds have entered clinical use. Successful clinical development of PEGylated pharmaceuticals requires accurate methods for the qualitative and quantitative analysis of intact PEG conjugates in biological fluids. In this article, we review assay methods that can be utilized for the detection and measurement of PEGylated pharmaceuticals in complex biological samples for determination of biodistribution and pharmacokinetic properties. In particular, we describe relevant colorimetric, chromatographic, radiolabeled, biological, and enzyme-linked immunosorbent assays for the pharmacokinetic study of PEGylated molecules.
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