Showing papers by "Inchan Kwon published in 2020"
TL;DR: An overview of the recent progress in the development of novel nanomaterials for the alleviation of hypoxic microenvironment is presented, with promising start in the early phase and expected to grow rapidly in the coming years.
129 citations
TL;DR: Results clearly show that by applying ABP-HSA strong interaction to injectable hydrogels and therapeutic protein, the concentration of the therapeutic protein can be maintained for a long period in vivo, prolonging its therapeutic effect.
16 citations
TL;DR: The platform described herein could be used to develop other therapeutic peptides with extended serum half-lives and had different in vitro biological activities and in vivo glucose-lowering effects, demonstrating the importance of site-specific HSA conjugation.
Abstract: The number of therapeutic peptides for human treatment is growing rapidly. However, their development faces two major issues: the poor yield of large peptides from conventional solid-phase synthesis, and the intrinsically short serum half-life of peptides. To address these issues, we investigated a platform for the production of a recombinant therapeutic peptide with an extended serum half-life involving the site-specific conjugation of human serum albumin (HSA). HSA has an exceptionally long serum half-life and can be used to extend the serum half-lives of therapeutic proteins and peptides. We used glucagon-like-peptide 1 (GLP-1) as a model peptide in the present study. A “clickable” non-natural amino acid—p-azido-l-phenylalanine (AzF)—was incorporated into three specific sites (V16, Y19, and F28) of a GLP-1 variant, followed by conjugation with HSA through strain-promoted azide–alkyne cycloaddition. All three HSA-conjugated GLP-1 variants (GLP1_16HSA, GLP1_19HSA, and GLP1_28HSA) exhibited comparable serum half-lives in vivo. However, the three GLP1_HSA variants had different in vitro biological activities and in vivo glucose-lowering effects, demonstrating the importance of site-specific HSA conjugation. The platform described herein could be used to develop other therapeutic peptides with extended serum half-lives.
13 citations
TL;DR: It is demonstrated that the intramolecular distance in the conjugate of Uox and FA govern the stable formation of FcRn/SA/FA-conjugated protein and serum half-life extension in vivo.
12 citations
06 Jul 2020
TL;DR: This work presents a new approach for co-immobilization of multiple enzymes for biosensing and synthetic applications that combines sorption, desorption, and aggregation for high-performance immobilization of enzymes.
Abstract: Enzyme immobilization is very important for diverse enzyme applications. Particularly, there is a growing need for co-immobilization of multiple enzymes for biosensing and synthetic applications. H...
9 citations
TL;DR: Test the hypothesis that a genetically encoded phenylalanine analogue containing a hydrogen-substituted tetrazine (frTet) would increase the IEDDA reaction rate, thereby allowing for successful bioconjugation in vivo, and demonstrated that the genetically encoded frTet allows an almost complete IEDda reaction in vivo upon addition of trans-cyclooctene, enabling temporal control of in vivo bioconjectation in a very high yield.
8 citations
TL;DR: Co‐overexpression of MjTyrRS‐Dopa and MjtRNATyrAUA increased the production yield of Dopa‐incorporated MAP foot protein type 3 (dfp‐3) by 57%.
Abstract: Mussel adhesive proteins (MAPs) have great potential as bioglues, particularly in wet conditions. Although in vivo residue-specific incorporation of 3,4-dihydroxyphenylalanine (Dopa) in tyrosine-auxotrophic Escherichia coli cells allows for production of Dopa-incorporated bioengineered MAPs (dMAPs), the low production yield hinders the practical application of dMAPs. This low production yield of dMAPs is due to low translational activity of a noncanonical amino acid, Dopa, in E. coli cells. Herein, to enhance the production yield of dMAPs, we investigated the coexpression of Dopa-recognizing tyrosyl-tRNA synthetases (TyrRSs). To use the Dopa-specific Methanococcus jannaschii TyrRS (MjTyrRS-Dopa), we altered the anticodon of tyrosyl-tRNA amber suppressor into AUA (MjtRNATyr AUA ) to recognize a tyrosine codon (AUA). Co-overexpression of MjTyrRS-Dopa and MjtRNATyr AUA increased the production yield of Dopa-incorporated MAP foot protein type 3 (dfp-3) by 57%. Similarly, overexpression of E. coli TyrRS (EcTyrRS) led to a 72% higher production yield of dfp-3. Even with coexpression of Dopa-recognizing TyrRSs, dfp-3 has a high Dopa incorporation yield (over 90%) compared to ones prepared without TyrRS coexpression.
7 citations
TL;DR: This work hypothesized that small proteins do not cause substantial competition for FcRn binding to albumin, resulting in the extended serum half-life, and investigated whether the intramolecular distance in FA-protein conjugate affects the FcCn binding with albumin and serumHalf-life using linkers with varying lengths.
Abstract: Conjugation of serum albumin or one of its ligands (such as fatty acid) has been an effective strategy to prolong the serum half-lives of drugs via neonatal Fc receptor (FcRn)–mediated recycling of albumin. So far, fatty acid (FA) has been effective in prolonging the serum half-lives for therapeutic peptides and small proteins, but not for large therapeutic proteins. Very recently, it was reported a large protein conjugated to FA competes with the binding of FcRn with serum albumin, leading to limited serum half-life extension, because primary FA binding sites in serum albumin partially overlap with FcRn binding sites. In order to prevent such competition, longer linkers between FA and the large proteins were required. Herein, we hypothesized that small proteins do not cause substantial competition for FcRn binding to albumin, resulting in the extended serum half-life. Using a small protein (28 kDa), we investigated whether the intramolecular distance in FA-protein conjugate affects the FcRn binding with albumin and serum half-life using linkers with varying lengths. Unlike with the FA-conjugated large protein, all FA-conjugated small proteins with different linkers exhibited comparable the FcRn binding to albumin and extended serum half-life.
6 citations
Patent•
03 Jun 2020
TL;DR: In this article, a glucagon-like peptide-1 derivative was conjugated with albumin through a linker to increase a half-life in vivo through conjugation.
Abstract: The present invention relates to a glucagon-like peptide-1 derivative conjugated with albumin and, more particularly, to a derivative having albumin conjugated with a certain position of the glucagon-like peptide-1 derivative through a linker, a use thereof and a method for preparing the same. According to the present invention, in case of a GLP-1 derivative having a GLP-1 mutant conjugated with human serum albumin through a linker, albumin is conjugated with a certain position of GLP-1, particularly 13Y or 22F, thereby minimizing a decrease in a medicinal effect due to a steric hindrance. In addition, the derivative according to the present invention increases a half-life in vivo through conjugation with albumin. Accordingly, the GLP-1 derivative having a GLP-1 mutant conjugated with human serum albumin through a linker may be useful in treating diabetics.