Development and evaluation of antibody-fluorophore-conjugates to detect endometrial tissue

Purpose Endometriosis is a chronic disease caused by ectopic endometrial tissue. Today’s diagnostic and therapeutic standard is the laparoscopic resection of these foci. However, identifying the pathological tissue can be challenging leading to incomplete resection and symptom persistence. Current imaging methods trying to address this issue lack specificity. We therefore developed antibody-fluorophore-conjugates (AFC) targeting antigens which are specific for endometrial cells and could potentially improve the detection specificity. In this study we aim to show that these conjugates are capable of specifically detecting endometrial cell lines in vitro.

Material and methods We generated recombinant antibody-fusion-proteins (AFP) containing the snap-tag, a self-labeling protein used for site-directed-conjugation with a fluorophore. These AFCs targeted four antigens known to be expressed by endometrial tissue: Epidermal Growth Factor Receptor (EGFR), Chondroitin sulfate proteoglycan 4 (CSPG4), Epithelial cell adhesion molecule (EpCAM) and Folate Receptor alpha (Fra). To show their detection capacity we incubated the conjugates with four endometrial cell lines (12z, 49z, T-HESC, Ishikawa) and analyzed the binding activity using FACS and fluorescence microscopy.

Results The conjugates were suitable for the specific detection of endometrial cell lines expressing the antigens EGFR, CSPG4, EpCAM and FRa. It was also possible to perform multiplex imaging by combining two different AFCs for the simultaneous detection of two antigens.

Conclusion AFCs generated by using the SNAP-tag-technology were able to detect endometrial cell lines by targeting endometrial antigens. However, the possible role as a diagnostic tool needs further study e.g. the ability of the conjugates to detect endometrial tissue samples.

Publikationsverlauf

Artikel online veröffentlicht:
11. Oktober 2022