Shashikant Ray, Sarah Jacques, Utibeabasi Ettah, Shaodong Dai, Hanmant K Gaikwad, Robert I Scheinman, Krishna M G Mallela, Dmitri Simberg
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引用次数: 0
Abstract
PEGylation is commonly utilized to modify nanoparticles, vaccines, therapeutic proteins, and biomaterials. The increased use of PEG-containing cosmetics and medicines results in anti-PEG immunoglobulins in humans, including IgG, IgM, and IgE types. Recent studies have shown that high-affinity antibodies against PEG can be isolated through immunization. These antibodies have been used in immunoassays and for decorating PEGylated nanoparticles with targeting ligands. However, conventional antibodies have limitations, such as their large size and potential stability issues. Here, we developed a protocol to isolate anti-PEG nanobodies (Nbs) from a yeast library. We prepared ∼60 nm fluorescently labeled PEGylated and non-PEGylated iron oxide nanoworms for negative and positive magnetic selection, followed by multiparameter flow cytometry sorting. The representative Nb clone exhibited highly specific binding to PEGylated liposomes and nanoparticles and was able to compete with commercial antibackbone PEG IgG but not with anti-methoxy PEG IgG for PEG binding. The Nb showed a micromolar dissociation constant, no aggregation, high thermal stability, and reversible structural stability. This work represents a proof-of-concept screening and isolation of anti-PEG Nbs.
期刊介绍:
Molecular Pharmaceutics publishes the results of original research that contributes significantly to the molecular mechanistic understanding of drug delivery and drug delivery systems. The journal encourages contributions describing research at the interface of drug discovery and drug development.
Scientific areas within the scope of the journal include physical and pharmaceutical chemistry, biochemistry and biophysics, molecular and cellular biology, and polymer and materials science as they relate to drug and drug delivery system efficacy. Mechanistic Drug Delivery and Drug Targeting research on modulating activity and efficacy of a drug or drug product is within the scope of Molecular Pharmaceutics. Theoretical and experimental peer-reviewed research articles, communications, reviews, and perspectives are welcomed.