@misc{9236655,
  abstract     = {{Recombinant protein development requires not only successful expression and purification, but also the recovery of stable and functionally active proteins. In this study, five recombinant proteins were used as case examples to examine key challenges in early-stage recombinant protein production and functional characterization. These included three oat seed lipase 1 variants, OL522, OL118, and OL183, and two enamel-related proteins, amelotin and amelogenin-mid. All constructs were expressed in Escherichia coli BL21(DE3) using a pET11 expression system. Protein expression, extraction, solubility, purification, refolding, and preliminary functional behavior were evaluated mainly by SDS-PAGE, immobilized metal affinity chromatography, fluorescence-based lipase activity assays, and turbidity-based mineralization analysis. 
The results showed that all five recombinant proteins were expressed and obtained after purification using their respective strategies. The three oat lipase variants showed enzymatic activity after refolding, although the activity was inconsistent and unstable. Amelotin showed calcium- and phosphate-dependent precipitation, indicating a possible mineralization-associated response.
Overall, the target recombinant proteins were successfully obtained, but reliable functional validation requires further investigation.}},
  author       = {{Bai, Yumei}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{From Expression to Function: Challenges in Recombinant Protein Development and Functional Characterization}},
  year         = {{2026}},
}