Journal: Frontiers in Immunology

Article Title: A rabbit anti-human CD38 antibody for eliminating daratumumab and isatuximab interference in immunohematology testing

doi: 10.3389/fimmu.2026.1726341

Figure Lengend Snippet: Expression, characterization, and functional evaluation of rabbit anti-hCD38 polyclonal antibody. (A) Purity and molecular weight of hCD38-His protein detected via SDS–PAGE and Coomassie brilliant blue staining. (B) Purity and molecular weight of rabbit anti-hCD38 pAb analyzed via SDS–PAGE and Coomassie staining. (C) Affinity of rabbit pAb to hCD38-His determined via ELISA (EC 50 = 86.93 ng/mL). (D) Results of indirect anti-human globulin tests in the presence (plus sign) or absence (minus sign) of DARA and pAb are shown. pAb-treated RBCs eliminated DARA-induced pan-agglutination in IAT. (E) Same as panel (D) , using ISA-spiked plasma. The results showed that treatment with pAb eliminated ISA-induced pan-agglutination in IAT. Rabbit pAb, rabbit anti-human CD38 polyclonal antibody; DARA, daratumumab; ISA, isatuximab; pAb, polyclonal antibody; RBCs, red blood cells; IAT, indirect antiglobulin test. A solid pellet at the bottom of the tubes indicates a negative result, and suspended particles (red cell agglutinates) within the gel matrix indicate a positive test result (either a 1+ or 2+ degree of agglutination).

Article Snippet: The human CD38 gene vector (Sino Biological, NP_001766 , Cat. HG10818-M, Beijing, China) was amplified using primers listed in .

Techniques: Expressing, Functional Assay, Molecular Weight, SDS Page, Staining, Enzyme-linked Immunosorbent Assay, Agglutination, Clinical Proteomics, Indirect Antiglobulin Test

Journal: Frontiers in Immunology

Article Title: A rabbit anti-human CD38 antibody for eliminating daratumumab and isatuximab interference in immunohematology testing

doi: 10.3389/fimmu.2026.1726341

Figure Lengend Snippet: Preparation, expression, and identification of rabbit anti-human CD38 monoclonal antibodies. (A) Flow cytometric sorting of single B cells; FITC/allophycocyanin (APC) (AF647) double-positive cells in AE gate selected. (B) Purity and molecular weight of rabbit mAbs were detected via staining with Coomassie brilliant blue. (C) ELISA analysis of rabbit mAb binding to hCD38-His. (D) ForteBio Octet determination of the affinities of rabbit mAbs toward hCD38-His. The binding affinity parameter KD was calculated, as reported using ForteBio Data Analysis Software 8.0 (Fremont, CA, USA). (E) Epitope competition assay with DARA by ForteBio. Green, A3; purple, D2. (F) Flow cytometry showing D2 and A3 binding to RBC-expressed CD38. Rabbit mAbs, rabbit anti-human CD38 monoclonal antibodies; DARA, daratumumab; RBC, red blood cell.

Article Snippet: The human CD38 gene vector (Sino Biological, NP_001766 , Cat. HG10818-M, Beijing, China) was amplified using primers listed in .

Techniques: Expressing, Bioprocessing, Molecular Weight, Staining, Enzyme-linked Immunosorbent Assay, Binding Assay, Software, Competitive Binding Assay, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: A rabbit anti-human CD38 antibody for eliminating daratumumab and isatuximab interference in immunohematology testing

doi: 10.3389/fimmu.2026.1726341

Figure Lengend Snippet: Elimination of DARA- and ISA-induced interference in IAT using rabbit anti-human CD38 monoclonal antibodies and optimization of D2 treatment conditions. Results of indirect anti-human globulin tests in the presence (plus sign) or absence (minus sign) of DARA/ISA and A3/D2 are shown. (A) A3-treated RBCs failed to eliminate DARA-induced interference in IAT. (B, C) D2-treated RBCs eliminated DARA- and ISA-induced pan-agglutination in IAT. (D) Tube 1: negative control. Tube 2: positive control for the DARA interference. Tubes 3–9: per μL of packed RBCs was treated with varying volumes of 1–30 μL D2 (1 mg/mL), and at least 3 μL of 1 mg/mL D2 was required to effectively eliminate DARA interference. (E) Per μL packed RBCs was treated with 3 μL of D2 at room temperature for varying durations (5–15 minutes). Optimization experiments demonstrated that an incubation time of 10 minutes at room temperature was sufficient to eliminate DARA interference in the presence of D2. DARA, daratumumab; ISA, isatuximab; IAT, indirect antiglobulin test; RBCs, red blood cells. A solid pellet at the bottom of the tubes indicates a negative result, and suspended particles (red cell agglutinates) within the gel matrix indicate a positive test result (either a 1+ or 2+ degree of agglutination).

Article Snippet: The human CD38 gene vector (Sino Biological, NP_001766 , Cat. HG10818-M, Beijing, China) was amplified using primers listed in .

Techniques: Bioprocessing, Agglutination, Negative Control, Positive Control, Incubation, Indirect Antiglobulin Test

Journal: Frontiers in Immunology

Article Title: A rabbit anti-human CD38 antibody for eliminating daratumumab and isatuximab interference in immunohematology testing

doi: 10.3389/fimmu.2026.1726341

Figure Lengend Snippet: Elimination of DARA- and ISA-induced interference in IAT using rabbit anti-human CD38 monoclonal antibodies and optimization of D2 treatment conditions. Results of indirect anti-human globulin tests in the presence (plus sign) or absence (minus sign) of DARA/ISA and A3/D2 are shown. (A) A3-treated RBCs failed to eliminate DARA-induced interference in IAT. (B, C) D2-treated RBCs eliminated DARA- and ISA-induced pan-agglutination in IAT. (D) Tube 1: negative control. Tube 2: positive control for the DARA interference. Tubes 3–9: per μL of packed RBCs was treated with varying volumes of 1–30 μL D2 (1 mg/mL), and at least 3 μL of 1 mg/mL D2 was required to effectively eliminate DARA interference. (E) Per μL packed RBCs was treated with 3 μL of D2 at room temperature for varying durations (5–15 minutes). Optimization experiments demonstrated that an incubation time of 10 minutes at room temperature was sufficient to eliminate DARA interference in the presence of D2. DARA, daratumumab; ISA, isatuximab; IAT, indirect antiglobulin test; RBCs, red blood cells. A solid pellet at the bottom of the tubes indicates a negative result, and suspended particles (red cell agglutinates) within the gel matrix indicate a positive test result (either a 1+ or 2+ degree of agglutination).

Article Snippet: The human CD38 gene vector (Sino Biological, NP_001766 , Cat. HG10818-M, Beijing, China) was amplified using primers listed in .

Techniques: Bioprocessing, Agglutination, Negative Control, Positive Control, Incubation, Indirect Antiglobulin Test

Journal: Frontiers in Immunology

Article Title: A rabbit anti-human CD38 antibody for eliminating daratumumab and isatuximab interference in immunohematology testing

doi: 10.3389/fimmu.2026.1726341

Figure Lengend Snippet: Stability of D2 and its efficacy in eliminating therapeutic anti-CD38 antibody interference. Results of indirect anti-human globulin (Coombs’) tests in the presence (plus sign) or absence (minus sign) of daratumumab and D2 are shown. (A) Distribution of anti-CD38 antibody titers (n = 49) and their association with clinical response status. (B) Agglutination scores comparing the efficacy of DTT and D2 in reducing therapeutic anti-CD38 antibody interference. (C) To evaluate the storage stability of D2, it was stored at 4°C, −20°C, and −80°C, and its ability to eliminate DARA interference was assessed via IAT after 1 month (D) , 3 months (E) , or 6 months. A solid pellet at the bottom of the tubes indicates a negative result, and suspended particles (red cell agglutinates) within the gel matrix indicate a positive test result (either a 1+ or 2+ degree of agglutination). DTT, dithiothreitol; DARA, daratumumab; IAT, indirect antiglobulin test.

Article Snippet: The human CD38 gene vector (Sino Biological, NP_001766 , Cat. HG10818-M, Beijing, China) was amplified using primers listed in .

Techniques: Agglutination, Indirect Antiglobulin Test

Journal: Nature Communications

Article Title: Enhancing CAR- and TCR-mediated targeting of cancer via an immune synapse-stabilizing receptor

doi: 10.1038/s41467-025-65897-4

Figure Lengend Snippet: A A schematic showing interaction between CLL1.CAR and CD38.SSR. CAR and SSR configurations are denoted on the right B Representative flow plots showing co-expression of CAR and SSRs on day7 post co-transduction (TD). SSR expression was measured by detection of a surrogate maker truncated NGFR. C Residual tumor counts after a 3-day culture of CD38+ Molm13 (AML) and CCRF-CEM (T-ALL) with non-transduced T cells (NT) or SSR T cells at an E:T ratio 1:4. Data represent 3 donors. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean+S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant). D Histograms showing CLL1 and CD38 levels in indicated cell lines. E Residual tumor counts and T-cell expansion after a 3-day co-culture of Molm13 and CCRF-CEM with CAR T cells at a E:T ratio 1:4. Data represent 6 donors from 3 independent experiments. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant). F Histograms showing CLL1 and CD38 levels in pre-sorted THP1 and CLL1-low THP1 post-sorting. G Residual tumor counts and fold T-cell expansion after 3 day culture at a E:T ratio 1:4. Data represent 6 donors from 2 independent experiments. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (*P < 0.05. ns, non-significant). H CD38 expression in the parental and knock-out line of Molm13. I Residual tumor counts and fold T-cell expansion after 3 day culture at a E:T ratio 1:4. Data represent 4 donors from 2 independent experiments. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant). (* P < 0.05, ** P < 0.01. ns, non-significant).

Article Snippet: To investigate LAT downstream signaling, 1 × 10 6 CAR.SSR T cells were loaded into a well of a 24-well plate pre-coated with 0.3 μg anti-CLL1 CAR antibody and 1 μg recombinant human CD38 (Sino Biological, Cat: 10818) for 30 min on ice, followed by stimulation at 37 °C in a water bath for indicated time-points.

Techniques: Expressing, Transduction, Co-Culture Assay, Knock-Out

Journal: Nature Communications

Article Title: Enhancing CAR- and TCR-mediated targeting of cancer via an immune synapse-stabilizing receptor

doi: 10.1038/s41467-025-65897-4

Figure Lengend Snippet: A Schematic illustration (left) and representative images (right) of calcium influx (green) in T cells (blue) at pre-contact or post-contact with Molm13 (red) over the indicated time course. B Calcium (Ca++) flux in T cells interacting with Molm13 over the indicated timepoints, and the mean area under the curve (AUC) for the initial 30 min post-contact. Each dot represents a single cell. Data include single cells (42 cells (NT), 34 cells (CAR), 30 cells (CAR + ΔSSR), 30 cells (CAR + SSR)) combined from 3 independent donors and experiments. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Dot plots show mean + S.E.M. (* P < 0.05, ** P < 0.01, ns, non-significant). C Western blots showing the detection of phospho-proteins and GAPDH harvested from CAR or CAR.SSR T cells stimulated with plate-coated anti-CLL1 CAR antibody and recombinant CD38 at indicated time points. Data were repeated in three independent experiments using 3 donors. MW, molecular weight. D Densitometry analysis of pPLCγ1, pERK1/2 and pNF-KB normalized to GAPDH. E Schematic of an SSR construct containing a point mutation Y132F in the LAT endo-domain, and flow plots (bottom) showing the co-expression of CAR and SSR or SSR with Y132F on day 6 post transduction. F Residual tumor counts (left) and T-cell expansion normalized to day 0 (right) in a 3 day co-culture with Molm13. Data represent two independent experiments with 4 donors. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (* P < 0.05, ns, non-significant).

Article Snippet: To investigate LAT downstream signaling, 1 × 10 6 CAR.SSR T cells were loaded into a well of a 24-well plate pre-coated with 0.3 μg anti-CLL1 CAR antibody and 1 μg recombinant human CD38 (Sino Biological, Cat: 10818) for 30 min on ice, followed by stimulation at 37 °C in a water bath for indicated time-points.

Techniques: Western Blot, Recombinant, Molecular Weight, Construct, Mutagenesis, Expressing, Transduction, Co-Culture Assay

Journal: Nature Communications

Article Title: Enhancing CAR- and TCR-mediated targeting of cancer via an immune synapse-stabilizing receptor

doi: 10.1038/s41467-025-65897-4

Figure Lengend Snippet: A Representative flow plots showing expression of CD38 and CLL1 in PBMCs including CD14+ monocytes, CD3 + T cells, CD3-CD56 + NK cells and CD19 + B cells. B Residual PBMCs after a 24-h co-culture with autologous T cells at a 1:4 E:T ratio. Data represent 4 donors. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (** P < 0.01. ns, non-significant). C Quantification of a burst forming unit-erythroid (BFU-E) and colony forming unit-granulocyte/macrophage (CFU-GM) from hematopoietic stem cells/progenitors (HSPC) after 5-h co-culture of indicated CAR T cells with CD34+ cord blood cells at a E:T ratio 10:1, and expanded for 12 days in semi-solid methylcellulose. Data represent 4 donors. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant). D Flow histograms show surface CLL1 and CD38 levels on AML blasts (CD33 + /CD34 + ) from the PBMCs collected from 2 patients (Pt1 and Pt2). E The residual AML blasts were detected at 24 h or 72 h after co-culture with CLL1 CAR T cells at a 1:1 E:T ratio. Data represent 3 independent donors. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons, or two-tailed, paired Student’s t test for CAR vs CAR + SSR in residual AML blasts from Pt2 at 72 h. Bar graphs show mean + S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant).

Article Snippet: To investigate LAT downstream signaling, 1 × 10 6 CAR.SSR T cells were loaded into a well of a 24-well plate pre-coated with 0.3 μg anti-CLL1 CAR antibody and 1 μg recombinant human CD38 (Sino Biological, Cat: 10818) for 30 min on ice, followed by stimulation at 37 °C in a water bath for indicated time-points.

Techniques: Expressing, Co-Culture Assay, Two Tailed Test

Journal: Nature Communications

Article Title: Enhancing CAR- and TCR-mediated targeting of cancer via an immune synapse-stabilizing receptor

doi: 10.1038/s41467-025-65897-4

Figure Lengend Snippet: A A schematic showing enhancement of cytotoxicity of survivin-specific TCR (Sur-TCR) by a CD38.SSR. B Representative flow plots showing co-expression of Sur-TCR and SSR on day 7 post-transduction (TD), and ( C ) expansion of TCR T cells derived from 3 HLA-A2- and 3 HLA-A2+ donors on day 10. Data represent 6 donors from two independent experiments. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (ns, non-significant). D Histograms showing intracellular survivin, surface HLA-A2 and CD38 levels in leukemia cell lines BV173, THP1 and TALL1. E Residual tumor counts after a 3-day co-culture with Sur-TCR T cells at a 1:4 E:T ratio. Data represent 6 donors from two independent experiments in co-culture with BV173 and THP1, and 3 donors from one independent experiment in co-culture with TALL1. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant). Percentages of ( F ) granzyme B/CD107a double positive T cells and ( G ) cytokine positive T cells after 4-h co-culture with indicated leukemia cells at 1:1 E:T ratio. Data represent 3 donors. P values were determined using one way ANOVA with Tukey’s correction for multiple comparisons. Bar graphs show mean + S.E.M. (* P < 0.05, ** P < 0.01, *** P < 0.001. ns, non-significant). H Schematic timeline of the mouse experiment. I Leukemia progression was monitored by bioluminescence. J Tumor burden AUC between day 0 and day 26. The data represent 5 mice in NT and 4 mice in for TCR, TCR.ΔSSR and TCR.SSR. P values were determined using one-way ANOVA with Dunnet’s correction for multiple comparisons compared to NT. Bar graphs show mean + S.E.M. (* P = 0.0394). K Overall mouse survival in the THP1 model (NT, n = 5, TCR, n = 5, TCR.ΔSSR, n = 5, TCR.SSR, n = 4). Statistical significance was determined by a log rank test. (* P = 0.0214).

Article Snippet: To investigate LAT downstream signaling, 1 × 10 6 CAR.SSR T cells were loaded into a well of a 24-well plate pre-coated with 0.3 μg anti-CLL1 CAR antibody and 1 μg recombinant human CD38 (Sino Biological, Cat: 10818) for 30 min on ice, followed by stimulation at 37 °C in a water bath for indicated time-points.

Techniques: Expressing, Transduction, Derivative Assay, Co-Culture Assay