Two Highly Specific Mouse Monoclonal Antibodies to the Putative C-Telopeptide of Human Collagen XIα1, a Cancer Biomarker

Background: Collagen XIα1, encoded by the COL11A1 gene, is a minor fibrillar collagen that is overexpressed in various human cancers, in which its presence correlates with tumor aggressiveness and progression. Methods: In this study, we developed two novel mouse monoclonal antibodies (mAbs), Anti-colXIα1 clone 3 and Anti-colXIα1 clone 9, that target the putative C-telopeptide of human collagen XIα1. The antibodies were raised to the RRHTEGMQA sequence, a unique nine-amino acid stretch within the putative C-telopeptide of human collagen XIα1. Results: Corresponding to nearly identical V(D)J gene segments and complementarity-determining regions (CDRs), the antibodies specifically bound the RRHTEGMQA epitope in ELISAs but did not react with the C-propeptide. This specificity was further confirmed with the purified Anti-colXIα1 clone 9 mAb, which demonstrated strong reactivity to recombinant proteins containing the RRHTEGMQA sequence in both ELISAs and Western blot assays. This sequence seems to behave as a linear B-cell neoepitope, in which the RRHT motif is crucial for epitope recognition. Otherwise, no immunodetections were observed either in cultures and lysates from the COL11A1-highly expressing A204 human cell line or on tissue sections from specimens of human pancreatic ductal adenocarcinoma (PDAC), with strong desmoplastic reactions, Conclusions: Lacking a precise knowledge of the characteristics of the putative C-telopeptide of human collagen XIα1, these antibodies could enhance our understanding of the processing of human procollagen XIα1 and contribute to a better characterization of the tumor microenvironment of COL11A1-expressing cancers.