Heat‐sterilizable antibody mimics designed on the cold shock protein scaffold from hyperthermophile Thermotoga maritima DOI Open Access
Hiroshi Amesaka,

M. Tachibana,

Mizuho Hara

et al.

Protein Science, Journal Year: 2024, Volume and Issue: 34(1)

Published: Dec. 26, 2024

Antibodies and antibody mimics are extensively used in the pharmaceutical industry, where stringent safety standards required. Implementing heat sterilization during or after manufacturing process could help prevent contamination by viruses bacteria. However, conventional antibodies not suitable for because they irreversibly denature at high temperatures. In this study, we focused on refolding property of cold shock protein from hyperthermophile Thermotoga maritima (TmCSP), which denatures elevated temperatures but regains its native structure upon re-cooling. We designed constructed a mutant library TmCSP amino acid residues three surface loops were diversified. From library, TmCSPs that bind to each eight target proteins selected phage yeast display methods. confirmed secondary binding affinity all mutants restored treatment followed cooling. Additionally, freeze-drying did impair their affinity. The crystal complex with target, esterase Alicyclobacillus acidocaldarius, revealed specific interactions between them. These results clearly demonstrate feasibility creating heat-sterilizable using as scaffold.

Language: Английский

Chemical Synthesis of Interleukin-6 for Mirror-Image VHH Discovery DOI

Keisuke Aoki

Springer theses, Journal Year: 2025, Volume and Issue: unknown, P. 85 - 114

Published: Jan. 1, 2025

Language: Английский

Citations

0
Synthetic Study of Full-Length Hepatitis B Virus Core Protein and Its Capsid Assembly DOI

Keisuke Aoki

Springer theses, Journal Year: 2025, Volume and Issue: unknown, P. 71 - 83

Published: Jan. 1, 2025

Language: Английский

Citations

0
Mirror-Image Human Serum Albumin Domain III as a Tool for Analyzing Site II-Dependent Molecular Recognition DOI

Naoya Iwamoto,

Takuma Kai,

Shinsuke Inuki

et al.

Bioconjugate Chemistry, Journal Year: 2024, Volume and Issue: 35(6), P. 816 - 825

Published: May 23, 2024

Human serum albumin (HSA) as a drug carrier can significantly improve the pharmacokinetic profiles of short-lived therapeutics. Conjugation albumin-binding moieties (ABMs) to therapeutic agents may prolong their half-life by promoting association with endogenous HSA. To discover new molecular class ABMs from mirror-image chemical space, preparation protocol for bioactive HSA domain III and its d-enantiomer (d-HSA III) was established. Structural functional analyses suggested that synthetic protein enantiomers exhibited structures stereoselective neonatal fragement crystallizable receptor (FcRn) recognition. Additionally, ligand-binding properties l-HSA were comparable those site II in native HSA, confirmed using II-selective fluorescent probes an esterase substrate. Synthetic d-HSA is attractive tool analyzing II-dependent recognition

Language: Английский

Citations

0
Chemical Synthesis of Interleukin-6 for Mirror-Image Screening DOI

Keisuke Aoki,

Kayuu Maeda,

Shinsuke Inuki

et al.

Bioconjugate Chemistry, Journal Year: 2024, Volume and Issue: 35(8), P. 1190 - 1199

Published: July 23, 2024

Interleukin-6 (IL-6), a multifunctional cytokine, is an attractive therapeutic target for immunological and inflammatory diseases. We investigated the chemical synthesis of IL-6 its enantiomer (d-IL-6) using sequential N-to-C native ligation strategy from six peptide segments. Solubilizing Trt-K

Language: Английский

Citations

0
Heat‐sterilizable antibody mimics designed on the cold shock protein scaffold from hyperthermophile Thermotoga maritima DOI Open Access
Hiroshi Amesaka,

M. Tachibana,

Mizuho Hara

et al.

Protein Science, Journal Year: 2024, Volume and Issue: 34(1)

Published: Dec. 26, 2024

Antibodies and antibody mimics are extensively used in the pharmaceutical industry, where stringent safety standards required. Implementing heat sterilization during or after manufacturing process could help prevent contamination by viruses bacteria. However, conventional antibodies not suitable for because they irreversibly denature at high temperatures. In this study, we focused on refolding property of cold shock protein from hyperthermophile Thermotoga maritima (TmCSP), which denatures elevated temperatures but regains its native structure upon re-cooling. We designed constructed a mutant library TmCSP amino acid residues three surface loops were diversified. From library, TmCSPs that bind to each eight target proteins selected phage yeast display methods. confirmed secondary binding affinity all mutants restored treatment followed cooling. Additionally, freeze-drying did impair their affinity. The crystal complex with target, esterase Alicyclobacillus acidocaldarius, revealed specific interactions between them. These results clearly demonstrate feasibility creating heat-sterilizable using as scaffold.

Language: Английский

Citations

0