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1. Intracellular Delivery of Proteins by Protein-Recognizing Nanoparticles

   https://doi.org/10.1021/acsami.4c18186  

   Ghosh, Avijit; Sharma, Mansi; Zhao, Yan (January 2025, ACS Applied Materials & Interfaces)
2. Materials for oral delivery of proteins and peptides

   https://doi.org/10.1038/s41578-019-0156-6  

   Brown, Tyler D.; Whitehead, Kathryn A.; Mitragotri, Samir (February 2020, Nature Reviews Materials)
3. Protein nanocage architectures for the delivery of therapeutic proteins

   https://doi.org/10.1016/j.cocis.2020.101395  

   Wang, Yang; Douglas, Trevor (February 2021, Current Opinion in Colloid & Interface Science)

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4. Microautophagy Mediates Vacuolar Delivery of Storage Proteins in Maize Aleurone Cells

   https://doi.org/10.3389/fpls.2022.833612  

   Ding, Xinxin; Zhang, Xiaoguo; Paez-Valencia, Julio; McLoughlin, Fionn; Reyes, Francisca C.; Morohashi, Kengo; Grotewold, Erich; Vierstra, Richard D.; Otegui, Marisa S. (February 2022, Frontiers in Plant Science)

   The molecular machinery orchestrating microautophagy, whereby eukaryotic cells sequester autophagic cargo by direct invagination of the vacuolar/lysosomal membrane, is still largely unknown, especially in plants. Here, we demonstrate microautophagy of storage proteins in the maize aleurone cells of the endosperm and analyzed proteins with potential regulatory roles in this process. Within the cereal endosperm, starchy endosperm cells accumulate storage proteins (mostly prolamins) and starch whereas the peripheral aleurone cells store oils, storage proteins, and specialized metabolites. Although both cell types synthesize prolamins, they employ different pathways for their subcellular trafficking. Starchy endosperm cells accumulate prolamins in protein bodies within the endoplasmic reticulum (ER), whereas aleurone cells deliver prolamins to vacuoles via an autophagic mechanism, which we show is by direct association of ER prolamin bodies with the tonoplast followed by engulfment via microautophagy. To identify candidate proteins regulating this process, we performed RNA-seq transcriptomic comparisons of aleurone and starchy endosperm tissues during seed development and proteomic analysis on tonoplast-enriched fractions of aleurone cells. From these datasets, we identified 10 candidate proteins with potential roles in membrane modification and/or microautophagy, including phospholipase-Dα5 and a possible EUL-like lectin. We found that both proteins increased the frequency of tonoplast invaginations when overexpressed in Arabidopsis leaf protoplasts and are highly enriched at the tonoplast surface surrounding ER protein bodies in maize aleurone cells, thus supporting their potential connections to microautophagy. Collectively, this candidate list now provides useful tools to study microautophagy in plants. 

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5. Multilayered Microneedles for Triphasic Controlled Delivery of Small Molecules and Proteins

   https://doi.org/10.1002/mabi.202300431  

   Wright, Nathaniel; Wu, Tim; Wang, Yadong (December 2023, Macromolecular Bioscience)

   Abstract Transdermal delivery is an attractive delivery method that increases bioavailability, is suitable for a wide variety of therapeutics, and offers stable delivery outcomes. However, many therapeutics are unable to readily cross the stratum corneum. Microneedles mechanically disrupt the cutaneous barrier to deliver small molecules, proteins, and vaccines. To date, microneedles have not been used in conjunction with coacervate, a liquid–liquid phase separation that protects unstable proteins. A three‐layer microneedle for the controlled release of three different molecules is designed. Through micromolding, microneedles are efficiently generated, which benefits product scalability. The microneedles have good mechanical integrity and effectively penetrate porcine skin ex vivo. The three layers, in the microneedles, release the cargo in a three‐phase manner. The released protein maintains its structure well. Moreover, layer thickness can be controlled by varying fabrication parameters. The microneedles can incorporate both small molecule drugs and protein therapeutics, thus promising uses in multi‐drug therapies through a single treatment. 

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