INTERNATIONAL CONGRESS OF
CancerGenetics
Targeted Detection of HER3-Expressing Cancer Cells via a Novel Bioluminescent Affibody
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Farnaz Karbasi,1,*
1. University of Isfahan
- Introduction: Breast cancer ranks as the second most frequently diagnosed cancer across both sexes, representing approximately 11.6 % of all cancer cases. However, among women, it holds the distinction of being the most commonly diagnosed cancer and the primary cause of cancer-related mortality. Breast cancer accounts for 24 % of new cancer cases and 15 % of cancer deaths in women, with lung and colorectal cancers following in terms of mortality rates. Pro jections indicate that by 2040, the incidence of new breast cancer cases is expected to rise by over 46 %. HER3 (ErbB3) is a member of the ErbB family of receptor tyrosine kinases that plays a critical role in the progression and drug resistance of several cancers, particularly breast cancer. Its overexpression has been associated with poor prognosis, making it an attractive target for diagnostic approaches. In this study, we designed and developed a novel recombinant biosensor based on a fusion construct encoding an anti-HER3 affibody (Z08699) linked to luciferase enzyme.
- Methods: The coding sequence was cloned into the pET-21, an expression vector and transformed into Escherichia coli BL21(DE3). Following IPTG induction, the fusion protein was successfully expressed and purified using Ni-NTA affinity chromatography. The new affiprobe can be used for detecting, studying and monitoring the HER3-expressing cells using a luminometer in fast and inexpensive assays.
- Results: These results suggest that this affibody fusion protein may serve as a promising tool for the sensitive and specific detection of HER3-expressing cells, potentially aiding in early cancer diagnosis and therapeutic monitoring.
- Conclusion: In this study, a novel affiprobe was designed to bind to HER3, and its binding to HER3 was examined in a breast cancer cell line. The N-terminus of the Z08699 affibody was fused to the C- terminus of CSRLuc8. This affiprobe was successfully expressed in Escherichia coli BL21 (DE3), and its functionality and specificity were investigated through cytobiochemistry studies. To our knowledge, this study presents the first probe designed for the luminometric detection of HER3, which may hold potential as a new molecular tool for in vitro diagnostics.
- Keywords: Recombinant protein, bioluminescent, Biosensor, Cancer detection, Luciferase
