In Silico Structural Dynamics and Downstream Bioprocess of Recombinant Human SUMO-IL-33
5 affiliations
- 1School of Pharmacy, University of Medicine and Pharmacy at Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam
- 2Department of Cell and Molecular Biology, Uppsala University, 75124 Uppsala, Sweden
- 3Department of Chemistry, Imperial College London, London SW7 2AZ, UK
- 4University of Health Sciences, Vietnam National University Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam
- 5Research Center for Discovery and Development of Healthcare Products, Vietnam National University Ho Chi Minh City, Ho Chi Minh City 700000, Vietnam
Abstract
Interleukin-33 (IL-33) is a therapeutic target in inflammatory diseases, yet inhibitor screening is hindered by high commercial protein costs. While expressing IL-33 as a SUMO fusion protein overcomes yield limitations in Escherichia coli, whether this intact chimera can directly serve as a screening surrogate remains unresolved. Here, we combined in silico simulations with empirical bioprocess optimization to establish a high-yield pipeline for native human IL-33 production. AI-based structure predictions (AlphaFold and RoseTTAFold) revealed distinct inter-domain dynamics mediated by a flexible linker. Molecular dynamics simulations of the compact RoseTTAFold model suggested potential steric occlusion near receptor-binding interfaces, whereas AlphaFold models exhibited extended conformations, highlighting model-dependent variations that warrant further structural verification. To restore accessibility, tag cleavage was optimized using 1.0% (mol/mol) SUMO protease at 4 °C for 1 h, achieving near-complete digestion. Sandwich ELISA demonstrated an 87.6% molar epitope recognition recovery for cleaved IL-33 versus only 12.0% for the intact fusion, providing empirical evidence of substantial functional epitope masking by the SUMO tag. Two-step IMAC purification yielded ~19% mass recovery (~32.6% molar recovery). These findings demonstrate that tag removal is operationally essential for IL-33 functional availability in screening applications.
Related
- JAK3-Mediated Structural Remodeling of β-Catenin Suppresses Epithelial-Mesenchymal Transition: Insights from In Silico Modeling.
- Nomenclature, classification, and in-silico structural analysis of MSP1a variants of Anaplasma marginale
- Preparation, Structural Elucidation, and In Silico Prebiotic Evaluation of Pectin Oligosaccharides from Apple Pectin
- In silico analysis and structural modeling of zebu cattle α-lactalbumin milk protein
- QSAR-Guided In Silico Design of Piperazinylanthranilamide Factor Xa Inhibitors: Applicability-Domain Assessment, ADMET Profiling, Molecular Docking, Molecular Dynamics, and MM/GBSA
BibTeX
@article{nguyen2026silico,
title = {{In Silico Structural Dynamics and Downstream Bioprocess of Recombinant Human SUMO-IL-33}},
author = {Kim-Hung Nguyen and Tan Thanh Mai and Thua-Phong Lam and Long-Hung Dinh Pham and Hoang-Han Do and Minh-Thanh Quach and Khac-Minh Thai and Minh-Quan Le and Quoc-Thai Nguyen},
journal = {Biomolecules},
year = {2026},
month = oct,
doi = {10.3390/biom16101451},
url = {https://www.mdpi.com/2218-273X/16/10/1451},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - In Silico Structural Dynamics and Downstream Bioprocess of Recombinant Human SUMO-IL-33
AU - Kim-Hung Nguyen
AU - Tan Thanh Mai
AU - Thua-Phong Lam
AU - Long-Hung Dinh Pham
AU - Hoang-Han Do
AU - Minh-Thanh Quach
AU - Khac-Minh Thai
AU - Minh-Quan Le
AU - Quoc-Thai Nguyen
PY - 2026
DA - 2026/10/01
JO - Biomolecules
DO - 10.3390/biom16101451
UR - https://www.mdpi.com/2218-273X/16/10/1451
AB - Interleukin-33 (IL-33) is a therapeutic target in inflammatory diseases, yet inhibitor screening is hindered by high commercial protein costs. While expressing IL-33 as a SUMO fusion protein overcomes yield limitations in Escherichia coli, whether this intact chimera can directly serve as a screening surrogate remains unresolved. Here, we combined in silico simulations with empirical bioprocess optimization to establish a high-yield pipeline for native human IL-33 production. AI-based structure predictions (AlphaFold and RoseTTAFold) revealed distinct inter-domain dynamics mediated by a flexible linker. Molecular dynamics simulations of the compact RoseTTAFold model suggested potential steric occlusion near receptor-binding interfaces, whereas AlphaFold models exhibited extended conformations, highlighting model-dependent variations that warrant further structural verification. To restore accessibility, tag cleavage was optimized using 1.0% (mol/mol) SUMO protease at 4 °C for 1 h, achieving near-complete digestion. Sandwich ELISA demonstrated an 87.6% molar epitope recognition recovery for cleaved IL-33 versus only 12.0% for the intact fusion, providing empirical evidence of substantial functional epitope masking by the SUMO tag. Two-step IMAC purification yielded ~19% mass recovery (~32.6% molar recovery). These findings demonstrate that tag removal is operationally essential for IL-33 functional availability in screening applications.
SN - 2218-273X
ER -