# Proteomic Analysis of Plant-Derived hIGF-1-Fc Reveals Proteome Abundance Changes Associated with Wound Healing and Cell Proliferation

## TL;DR

This study explores how plant-made hIGF-1-Fc affects cell growth and healing by analyzing changes in proteins in mouse fibroblasts.

## Contribution

The first proteome-level analysis of plant-derived hIGF-1-Fc effects on mammalian cells is presented.

## Key findings

- hIGF-1-Fc promotes cancer cell migration and fibroblast proliferation.
- Proteomic changes include increased cytoskeletal proteins and metabolic enzymes.
- Gene ontology shows modulation of ribosome biogenesis and carbon metabolism.

## Abstract

Background: Human insulin-like growth factor 1 (hIGF-1) plays a key role in cell proliferation and tissue repair. While plant expression systems offer a cost-effective and scalable alternative for recombinant protein production, the molecular effects of plant-derived hIGF-1 on mammalian cells remain largely unexplored. Methods: In this study, a recombinant fusion protein of hIGF-1 with human Fc (hIGF-1-Fc) was transiently expressed in Nicotiana benthamiana using the geminiviral pBYR2e system and purified by Protein A affinity chromatography. SDS-PAGE and Western blotting confirmed the predicted molecular weight, and LC-MS identified N-glycosylation at the Fc N229 site with plant-type glycans such as GnMXF, GnGnXF, and MMXF. Bioactivity was evaluated using MCF-7 cell proliferation and NIH3T3 wound healing assays. Label-free quantitative proteomics was performed on NIH3T3 fibroblasts to assess molecular changes. Results: hIGF-1 Fc significantly promoted cancer cell migration and fibroblast proliferation. Proteomic profiling revealed an abundance of cytoskeletal proteins such as actin and tubulin and metabolic enzymes related to energy production. Gene ontology and pathway enrichment analyses indicated significant modulation of ribosome biogenesis and carbon metabolism. Conclusions: This study presents the first proteome-level investigation of plant-produced hIGF-1-Fc in mouse fibroblasts and reveals its impact on cytoskeletal organization and metabolic pathways involved in proliferation and wound healing.