Recombinant Expression of a Ready-to-Use EGF Variant Equipped With a Single Conjugation Site for Click-Chemistry.

ABSTRACT: The epidermal growth factor (EGF) receptor is commonly targeted in cancer therapy because it is overexpressed in many malignant cells. However, a general problem is to couple the targeting moieties and the drug molecules in a way that results in a homogeneous product. Here, we overcome this issue by engineering a variant of EGF with a single conjugation site for coupling virtually any payload. The recombinant EGF variant K-EGF^RR was expressed in E. coli Rosetta with a 4-6 mg/L yield. To confirm the accessibility of the introduced functional group, the ligand was equipped with a sulfo-cyanine dye with a loading of 0.65 dye per ligand, which enables tracking in vitro. The kinetics and affinity of ligand-receptor interaction were evaluated by enzyme-linked immunosorbent assa

SUBMITTER: Krass M 

PROVIDER: S-EPMC11913717 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature