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. 2023 May 7;24(9):8411. doi: 10.3390/ijms24098411

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Anti-human CD44 mAbs production. (A) The CD44 structure. CD44s mRNA is assembled by the first five (1 to 5) and the last five (16 to 20) exons and translates CD44s. CD44v mRNAs are generated by the alternative splicing of variant exons and translate multiple CD44v isoforms, such as CD44v3-10, CD44v4-10, CD44v6-10, and CD44v8-10. (B) BALB/c mice were intraperitoneally immunized with CHO/CD44v3–10 cells. (C) The hybridomas were produced via fusion of the splenocytes and P3U1 cells. (D) The flow cytometry-mediated screening was conducted by using parental CHO-K1 and CHO/CD44v3–10 cells. (E) After cloning and additional screening, a clone C₄₄Mab-6 (IgG₁, kappa) was established. Finally, the binding epitope was determined via enzyme-linked immunosorbent assay (ELISA) by using peptides, which cover the extracellular domain of CD44v3–10.