Expression, purification, and function of baculovirus infected insect cells‐derived hookworm AIP‐1 and AIP‐2 proteins

Abstract Anti‐inflammatory protein (AIP)‐1 and AIP‐2, identified as parasite excretory/secretory (ES) proteins, play a crucial role in promoting the survival of the parasite and evading the host immunological response. Both proteins inhibit inflammatory reactions, induce apoptosis in effector cells, and influence the phenotype of the immune response. Numerous parasite‐derived proteins have shown promise as therapeutic targets for inflammatory and allergic diseases. Despite this, the precise biological roles, and molecular characteristics of many such proteins remain unclear. In this study, AIP‐1 and AIP‐2 were produced in the baculovirus‐insect cell expression system (BEVS). The multiplicity of infection (MOI) and the duration of infection conditions for AIP‐1 and AIP‐2 protein expression were successfully optimized to induce the highest expression levels of AIP‐1 and AIP‐2 proteins in insect cells. The insect cell‐derived AIP‐1 and AIP‐2 exhibited inhibitory functions against human matrix metalloproteinases (MMPs). This study demonstrates that insect cell‐derived AIP‐1 and AIP‐2 have the potential as therapeutic proteins for MMP‐TIMP (Metalloprotease‐Tissue inhibitor of metalloprotease) axis related disease.