Journal: Molecular Therapy Oncology
Article Title: Chimeric antigen receptor T cell targeting CD44E variant in HCC holds therapeutic potential
doi: 10.1016/j.omton.2026.201181
Figure Lengend Snippet: CD44E-specific antibody screening and selection (A) The titers of anti-CD44E antibodies produced by hybridoma colonies were determined by ELISA. Anti-pan CD44 antibodies were selected as controls. (B) Binding of anti-CD44E-positive hybridoma supernatants to CD44E + cells was analyzed by flow cytometry. (C) Binding of anti-pan CD44-positive hybridoma supernatants to CD44E + cells was analyzed by flow cytometry. (D) Serial dilutions of purified anti-CD44E antibody (from S431 hybridoma colony) were added to ELISA plates coated with hCD44E-Fc and hCD44-Fc proteins to measure their binding activity and determine EC 50 values. (E) Surface plasmon resonance (SPR) analysis of S431 antibody binding to human CD44 isoforms. Real-time binding sensorgrams showing the interaction of serially diluted human CD44E-hFc (left) and human CD44-hFc (right) with immobilized S431 antibody. The binding response is indicated in resonance units (RU). (F) Flow cytometric analysis of antibody-binding specificity. The binding of the anti-CD44E antibody (S431 clone) was evaluated in HKCI-C1 cells overexpressing CD44E or CD44S (left), while the binding of anti-pan CD44 antibody (S129 clone) was assessed in the same cells (right). Human IgG (hIgG) was used as an isotype control. (G) Immunohistochemical (IHC) staining of CD44E in primary human liver tumor tissues (2 representative cases). Scale bars, 200 μm. (H) Representative immunofluorescent staining detecting cell-surface expression of CD44E protein in HKCI-11 cells. Scale bars, 100 μm.
Article Snippet: For CD44E expression, TaqMan assay for CD44E (Hs01081475_m1) and internal control 18S rRNA (Hs99999901_s1) were purchased from Applied Biosystems.
Techniques: Selection, Produced, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Purification, Activity Assay, SPR Assay, Control, Immunohistochemical staining, Immunohistochemistry, Staining, Expressing