A fusion protein and a complex containing same, capable of being used for the intracellular delivery of cargo molecules. The fusion protein and complex can implement the efficient release of cargo molecules from endocytic vesicles, thereby significantly improving the cytoplasmic delivery efficiency of the cargo molecules. One cargo molecules can be obtained in cytoplasms, they can exert any function related thereto. The fusion protein and complex provide effective means for affecting biological mechanisms and pathways of cells, and can be used in various fields such as research, treatment, and diagnosis.

The invention relates generally to methods for preparing recombinant nucleosomes. In particular, the invention relates to methods for ligating a histone peptide onto a fully assembled recombinant nucleosome. The invention further relates to modified core histone proteins, histone peptides to be ligated to the modified core histone proteins, and fully assembled recombinant nucleosomes and libraries of recombinant nucleosomes prepared by the methods of the invention.

This invention relates to a method of producing a circularised form of a target protein from a fusion protein and to a fusion protein capable of producing a circularised form of a target protein. The fusion protein can comprise: the target protein; at least one circularisation site adjacent the target protein; and an enzyme domain capable of interacting with the at least one circularisation site and circularising the target protein. The target protein can be a membrane scaffold protein or a cyclotide such as SFTI, Vc1.1, Kalata B1 or MCOTI-II.

A recombinant fusion protein includes a receptor associated protein 1 (RAP1), a cluster of differentiation 28 (CD28)-Pseudomonas exotoxin A translocation domain (PEt) fusion polypeptide, a legumain protein and a target peptide. The RAP1 is located at the N-terminus of the recombinant fusion protein. The CD28-PEt fusion polypeptide is located at the C-terminus of the RAP1. The legumain protein is located at the C-terminus of the CD28-PEt fusion polypeptide. The target peptide is located at the C-terminus of the legumain protein. In another embodiment of the present disclosure, an immunogenic composition is provided. The immunogenic composition including the recombinant fusion protein and an adjuvant is used for inducing specific immune responses in a subject with cancer, whereby the risk of cancer metastasis and recurrence for the subject may be successfully reduced.

The invention relates generally to methods for preparing recombinant nucleosomes. In particular, the invention relates to methods for ligating a histone peptide onto a fully assembled recombinant nucleosome. The invention further relates to modified core histone proteins, histone peptides to be ligated to the modified core histone proteins, and fully assembled recombinant nucleosomes and libraries of recombinant nucleosomes prepared by the methods of the invention

The present invention relates to methods that employ enzymes having Asx-specific ligase and cyclase activity, namely butelase-1, VyPAL2 and OaAEPI b, as a means for engineering novel (poly) peptide theranostics. The differential substrate specificities and differential optimal pH of the Asx-specific ligase and cyclase are used to provide sufficient orthogonality for a tandem ligation and cyclization of proteins. Also encompassed are the corresponding uses.

The present disclosure relates generally to generation of a recombinant enzyme with cyclization activity and its use for generating cyclic peptides as well as linear peptide conjugates.

The present invention lies in the technical field of enzymatic (poly)peptide ligation and specifically relates to methods that allow the ligation of (poly)peptides and oligonucleotides. The methods comprise providing at least one cargo molecule modified with a peptide tag and at least one poly(peptide) to be ligated to the cargo molecule, wherein the peptide tag and/or the (poly)peptide comprises a ligation motif for a peptide ligase, preferably sortase and peptidyl asparaginyl ligases (PALs), such as butelase-1, VyPAL2 or OaAEPI b. The invention also relates to the resulting conjugates and the corresponding uses.

Disclosed herein are methods for harvesting a recombinant protein comprising an asparaginyl endopeptidase (AEP) cleavage site, as well as methods for inhibiting cleavage of a recombinant protein comprising an AEP cleavage site, during a biomanufacturing process.