Encapsulating Proteins in Nanoparticles: Batch by Batch or One by One.

Encapsulation of proteins in nanoparticles (NPs) can greatly improve the properties of proteins such as their stability against denaturation and degradation by proteases, and branches out the applications of natural proteins from their intrinsic localizations and functions in living organisms for biomedical and industrial applications. We recently developed several methods to armor proteins in NPs with sizes from nanometers up to >100nm, batch by batch or one by one, covalently or noncovalently, for a wide range of applications from biocatalysis to bioimaging and drug delivery. In this chapter, we provide detailed protocols on these methods. Key steps of specific protocols are explained with particular examples to help other laboratories to adopt and modify these methods for their own purposes. The advantages and disadvantages of each method are summarized, and guidelines for choosing the right method for a given application, as well as the current challenges and future directions of this field, are discussed.