[Mild solubilization, purification, and immunoreactivity characterization of VP1 inclusion body protein of serotype O foot-and-mouth disease virus based on sodium lauroyl sarcosinate].

Authors

Lei Fan, Jianpeng Liang, Jieshan Lin, Linlin Cai, Tao Ren

Published in

Sheng wu gong cheng xue bao = Chinese journal of biotechnology. Volume 42. Issue 8. Pages 3541-3549. Aug 25, 2026.

Abstract

To efficiently prepare the capsid protein VP1 of serotype O foot-and-mouth disease virus (FMDV) with favorable immunoreactivity and to address the challenges that VP1 is prone to form inclusion bodies in the Escherichia coli expression system and is difficult to refold, this study developed a novel process based on sodium lauroyl sarcosinate (LSN) for the mild solubilization and chromatographic purification of FMDV VP1 inclusion body protein. First, the recombinant SUMO-VP1 protein was expressed in E. coli by induction, and Western blotting results revealed that the recombinant SUMO-VP1 protein was expressed at a high level; however, even in the presence of the SUMO solubility-enhancing tag, a large proportion of the protein still existed in the form of insoluble inclusion bodies, demonstrating the extremely hydrophobic folding propensity of the VP1 protein. Second, different concentrations of LSN were used to screen for mild solubilization conditions for the inclusion bodies. SDS-PAGE and grayscale analysis indicated that the solubilization rate exhibited a significant dose-dependent increase with rising LSN concentration, and 1.0% LSN was identified as the optimal working concentration for mild solubilization, at which the relative solubilization rate of the target protein reached as high as 92%. Subsequently, to avoid steric hindrance interference from the macromolecular tag, this 1.0% LSN solubilization process was applied to the extraction of SUMO-tag-free recombinant His-VP1 inclusion bodies, and the solubilized recombinant His-VP1 protein was purified using Ni²⁺-TED affinity chromatography, which successfully removed background contaminating proteins and yielded recombinant His-VP1 protein with a purity over 90%. Finally, the purified and desalted recombinant His-VP1 protein was used as the coating antigen in an indirect ELISA. The results demonstrated that the recombinant protein could be recognized by specific antibodies against VP1 of serotype O FMDV, and the binding curve exhibited a typical smooth sigmoidal shape and a significant dose-dependent effect, confirming that this process did not compromise the protein structure and fully preserved its native immunoreactivity. In conclusion, the LSN-based mild solubilization and chromatographic purification process developed in this study successfully circumvents the destructive effects and complex refolding steps associated with conventional strong denaturants, and the prepared recombinant His-VP1 protein possesses the potential to serve as a high-quality serological diagnostic antigen. This mild solubilization process may also provide a technical reference for the preparation of other highly hydrophobic and poorly soluble recombinant antigens.

PMID:
42638054
Bibliographic data and abstract were imported from PubMed on 25 Aug 2026.

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