mouse anti human cd22 antibody  (OriGene)

Journal: Molecular Therapy Oncology

Article Title: A measles virus encoding a CD19/CD3 bispecific T cell engager shows enhanced preclinical anti-BCP-ALL efficacy without significant toxicity

doi: 10.1016/j.omton.2026.201127

Figure Lengend Snippet: MV-Blina therapy significantly prolongs the survival of ALL bearing mice (A) Treatment scheme. Two PDX models were established in NSG mice by i.v. injection to induce ALL. At a leukemic load of approximately 35%, mice received treatment i.v. with MV (2.5 × 10 5 TCID 50 /g) or control. Two days after infection, mice were injected with healthy human PBMCs. (B) Superior survival of PDX #2 with MV-Blina compared to MV-Edm. PDX #1 and #2 were treated either with MV-Blina and PBMC ( n = 10) or MV-Edm and PBMC ( n = 10) or control with PBMC only ( n = 10) and observed for a maximum of 70 days. (C) MV-Blina treatment decreases leukemic load. Leukemic blasts (msCD45 − huCD19 + huCD45 dim ) and human PBMC (msCD45 − huCD19 + huCD45 bright ) in peripheral blood were monitored by flow cytometry at the indicated time points. (D) Marked reduction of spleen weight in PDX upon MV-Blina treatment. At the time of death, a complete necropsy was performed, and spleen weight was determined. (E) Significant reduction of leukemic blasts by MV-Blina in ALL compartments. At the time of death, leukemic blasts (msCD45 − huCD19 + huCD45 dim ) were detected by flow cytometry in bone marrow, spleen, and meninges. Replication of secBlina was detected by qRT-PCR and calculated by the 2 −ΔΔCt method, displaying the fold change relative to huActin and huGAPDH. (F) MV-Blina reduces leukemic blasts in the spleen by decreasing proliferation and increasing apoptosis. Spleens of untreated mice at therapy start (d0, n = 4) and treated mice (MV-Blina and PBMC, n = 10; MV-Edm and PBMC, n = 9; PBMC-only control, n = 9) were formalin-fixed, paraffin-embedded, and subsequently stained for H&E, CD22, Ki67, or cleaved Caspase-3 (CC3). Positive area was determined using ImageJ. Scale bars 50 μm. Statistical analysis was performed using Mantel-Cox log rank test (B), one-way ANOVA with Tukey’s multiple comparisons test (C-F). ns, not significant; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; and ∗∗∗∗, p < 0.0001.

Article Snippet: Immunohistochemistry for CD22 was performed using a mouse anti-human CD22 antibody (clone OTI1F2, 1:75, TA506404, Origene, Rockville, MD), for Ki67 using a mouse anti-human Ki67 antibody (clone MIB-1, 1:150, M7240, Dako, Waldbronn, Germany) and for cleaved caspase-3 (CC3) using a rabbit-anti human active caspase-3 antibody (1:200, ab13847, Abcam, Cambridge, UK).

Techniques: Injection, Control, Infection, Flow Cytometry, Quantitative RT-PCR, Formalin-fixed Paraffin-Embedded, Staining

Journal: Biophysical Reports

Article Title: NEU1 and NEU3 enzymes alter CD22 organization on B cells

doi: 10.1016/j.bpr.2022.100064

Figure Lengend Snippet: Cluster size of CD22 after treatment with cytoskeletal disruptors. Raji cells were treated with cytochalasin D or latrunculin A at 37 °C for 30 min. ( A ) Cells were then fixed and stained with mouse anti-CD22 and anti-mouse IgG-AF647 and imaged using confocal microscopy. ( B ) Data shown are average from 30 cells among 3 biological replicates. n values for conditions were 100 to 200 clusters. Cells were analyzed using ImageJ and are shown as beanplots . Comparison analyses were done using a one-way ANOVA followed by Dunnett’s t -test (∗∗∗∗p < 0.0001; ∗p < 0.05).

Article Snippet: Samples were treated with 1 μL/mL mouse anti-human IgM (clone IM260, Abcam, Cambridge, UK, cat# ab200541) or mouse anti-human CD22 (clone HIB22, BD Pharmingen, San Diego, CA, USA, cat# 555423) at 4 °C overnight and stained with goat anti-mouse IgG (polyclonal, Sigma-Aldrich, Burlington, MA, USA, cat# M8642) conjugated with Alexa Flour 647 (AF647) at room temperature for 1 h. The loading of the fluorophores was approximately 2 dyes/protein.

Techniques: Staining, Confocal Microscopy

Journal: Biophysical Reports

Article Title: NEU1 and NEU3 enzymes alter CD22 organization on B cells

doi: 10.1016/j.bpr.2022.100064

Figure Lengend Snippet: Lateral mobility of CD22 after treatment with cytoskeletal disruptors. Raji cells were treated at 37 °C for 30 min. Lateral mobility was analyzed using single-particle tracking with total internal fluorescence microscopy videos recorded at 10 frames per s for 10 s . Diffusion coefficients are given as log(D), where D is in units of × 10 −10 [cm 2 s −1 ] or × 10 −2 [μm 2 s −1 ]. 150 cells among 3 biological replicates were analyzed, and values were compared with control using one-way ANOVA followed by Dunnett’s test (∗∗∗∗p < 0.0001). Beanplots were generated using R software. Individual data points are represented by short white lines, a solid black line indicates the average for each condition, and the dotted line represents an average across all populations.

Article Snippet: Samples were treated with 1 μL/mL mouse anti-human IgM (clone IM260, Abcam, Cambridge, UK, cat# ab200541) or mouse anti-human CD22 (clone HIB22, BD Pharmingen, San Diego, CA, USA, cat# 555423) at 4 °C overnight and stained with goat anti-mouse IgG (polyclonal, Sigma-Aldrich, Burlington, MA, USA, cat# M8642) conjugated with Alexa Flour 647 (AF647) at room temperature for 1 h. The loading of the fluorophores was approximately 2 dyes/protein.

Techniques: Single-particle Tracking, Fluorescence, Microscopy, Diffusion-based Assay, Generated, Software

Journal: Biophysical Reports

Article Title: NEU1 and NEU3 enzymes alter CD22 organization on B cells

doi: 10.1016/j.bpr.2022.100064

Figure Lengend Snippet: CD22 cluster size is altered by NEU1 and NEU3 knockdown. Raji cells were transfected with siRNA targeting Neu1 or Neu3 using electroporation and grown for 24 h. ( A ) Western blots of transfected Raji cells. ( B ) Quantification of Western blots confirmed reduced expression of NEU1 and NEU3. ( C and D ) After transfection, Raji cells were fixed and stained with mouse anti-CD22 and anti-mouse IgG-AF647 and imaged using confocal microscopy ( C ) to determine the ( D ) cluster size of CD22. Data shown are average from 30 cells among 3 biological replicates. n values for each condition were between 100 and 200 clusters. Cells were analyzed using ImageJ and are shown as beanplots . Comparison analyses were done using one-way ANOVA followed by Dunnett’s t -test (∗∗p < 0.01; ∗p < 0.05).

Article Snippet: Samples were treated with 1 μL/mL mouse anti-human IgM (clone IM260, Abcam, Cambridge, UK, cat# ab200541) or mouse anti-human CD22 (clone HIB22, BD Pharmingen, San Diego, CA, USA, cat# 555423) at 4 °C overnight and stained with goat anti-mouse IgG (polyclonal, Sigma-Aldrich, Burlington, MA, USA, cat# M8642) conjugated with Alexa Flour 647 (AF647) at room temperature for 1 h. The loading of the fluorophores was approximately 2 dyes/protein.

Techniques: Transfection, Electroporation, Western Blot, Expressing, Staining, Confocal Microscopy

Journal: Biophysical Reports

Article Title: NEU1 and NEU3 enzymes alter CD22 organization on B cells

doi: 10.1016/j.bpr.2022.100064

Figure Lengend Snippet: CD22 expression after NEU1 and NEU3 knockdown. Raji cells were transfected with siRNA targeting Neu1 , Neu3 , or a scrambled control using electroporation. Cells were allowed to grow for 24 h, collected, and stained using mouse anti-CD22 primary antibody and goat anti-IgG secondary antibody. ( A and B ) A histogram of the fluorescence channel is shown ( A ), and quantitation of these data found no changes in expression for the siRNA treatments ( B ).

Article Snippet: Samples were treated with 1 μL/mL mouse anti-human IgM (clone IM260, Abcam, Cambridge, UK, cat# ab200541) or mouse anti-human CD22 (clone HIB22, BD Pharmingen, San Diego, CA, USA, cat# 555423) at 4 °C overnight and stained with goat anti-mouse IgG (polyclonal, Sigma-Aldrich, Burlington, MA, USA, cat# M8642) conjugated with Alexa Flour 647 (AF647) at room temperature for 1 h. The loading of the fluorophores was approximately 2 dyes/protein.

Techniques: Expressing, Transfection, Electroporation, Staining, Fluorescence, Quantitation Assay

Journal: Biophysical Reports

Article Title: NEU1 and NEU3 enzymes alter CD22 organization on B cells

doi: 10.1016/j.bpr.2022.100064

Figure Lengend Snippet: Lateral mobility of CD22 after treatment with NEU enzymes. Raji cells were treated at 37 °C for 30 min. Lateral mobility was analyzed using single-particle tracking with total internal fluorescence microscopy videos recorded at 10 frames per s for 10 s . Diffusion coefficients are given as log(D), where D is in units of × 10 −10 [cm 2 s −1 ] or × 10 −2 [μm 2 s −1 ]. Data shown are from 150 cells among 3 biological replicates. Data were analyzed and compared with control using a one-way ANOVA followed by Dunnett’s test (∗∗∗p < 0.005; ∗p < 0.05).

Article Snippet: Samples were treated with 1 μL/mL mouse anti-human IgM (clone IM260, Abcam, Cambridge, UK, cat# ab200541) or mouse anti-human CD22 (clone HIB22, BD Pharmingen, San Diego, CA, USA, cat# 555423) at 4 °C overnight and stained with goat anti-mouse IgG (polyclonal, Sigma-Aldrich, Burlington, MA, USA, cat# M8642) conjugated with Alexa Flour 647 (AF647) at room temperature for 1 h. The loading of the fluorophores was approximately 2 dyes/protein.

Techniques: Single-particle Tracking, Fluorescence, Microscopy, Diffusion-based Assay

Journal: Biophysical Reports

Article Title: NEU1 and NEU3 enzymes alter CD22 organization on B cells

doi: 10.1016/j.bpr.2022.100064

Figure Lengend Snippet: Model of changes in CD22 organization. ( A ) The size of the CD22 cluster formed is a result of interactions between CD22 and sialylated ligands, such as CD45, which can act as a bridge to homoclusters. A change in the stoichiometric ratio will shift the amount of complex (e.g., excess of the sialoside ligand or CD22). Increased NEU activity could reduce the total number of sialylated ligands available for CD22. ( B ) Cytoskeletal disruption allows microclusters of sialoglycoproteins to reorganize and act as a bridge to generate larger CD22 clusters. More extensive cytoskeletal disruption alters microclusters, leading to more diffuse clusters of both components. ( C ) A top-down representation of changes to cluster size due to cytoskeletal disruption.

Article Snippet: Samples were treated with 1 μL/mL mouse anti-human IgM (clone IM260, Abcam, Cambridge, UK, cat# ab200541) or mouse anti-human CD22 (clone HIB22, BD Pharmingen, San Diego, CA, USA, cat# 555423) at 4 °C overnight and stained with goat anti-mouse IgG (polyclonal, Sigma-Aldrich, Burlington, MA, USA, cat# M8642) conjugated with Alexa Flour 647 (AF647) at room temperature for 1 h. The loading of the fluorophores was approximately 2 dyes/protein.

Techniques: Activity Assay

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Relative fold expression of CD22 mRNA as compared to GAPDH in the cell lines included in the study, as measured by qRT- PCR a

Article Snippet: Mouse anti-human CD22 MAbs (HB22-7 or RFB4) and chimeric anti-human CD20 MAb, RituximabTM (Biogen Idec, Inc., San Diego, CA), were added in a final volume of 200 μL at concentrations ranging from 5 to 50 μg/mL (3.33-33.3 × 10 −8 M).

Techniques: Expressing, Quantitative RT-PCR, Gene Expression

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Representative PCR amplification of three distinct CD22 transcript regions using cDNA generated from the designated cell lines (1- Daudi; 2 - Jurkat; 3 - HCC827; 4 - H1355; 5 - H1975; 6 - A549; 7 - Calu-1; 8 - H1650; 9 - H727; 10 - dH2O). Daudi and Jurkat cells served as positive and negative controls, respectively. After 30 cycles of amplification, faint bands corresponding to CD22 transcripts were observed for Calu-1, H727, HCC827, H1650 and H1975 relative to the control (Daudi cells). Similar results were obtained in three or more experiments.

Article Snippet: Mouse anti-human CD22 MAbs (HB22-7 or RFB4) and chimeric anti-human CD20 MAb, RituximabTM (Biogen Idec, Inc., San Diego, CA), were added in a final volume of 200 μL at concentrations ranging from 5 to 50 μg/mL (3.33-33.3 × 10 −8 M).

Techniques: Amplification, Generated, Control

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: A. Flow cytometric analysis. One million cells from each cell line were incubated with 25 µg/mL of either a mouse isotype control antibody (MPC-11) or mouse antihuman CD22 (clone HB22.7) MAb. After washing out the unbound primary antibody, a FITC-labeled secondary goat anti-human IgG antibody was used. Cells were analyzed in FL-1 using a BD FACSCalibur. Red line – cells without antibodies; green line – cells plus the isotype control antibody and secondary antibody; blue line – cells plus the anti-CD22 antibody and secondary antibody. This is one representative out of 4–6 independent experiments. B. WB analysis. Equal amounts (25 µg) of protein from cell lysates were subjected to 8% SDS-PAGE followed by a WB with a rabbit anti-human CD22 antibody (H-221 clone). Upper panel - CD22 expression; lower panel - β-actin expression (loading control): 1 – Daudi; 2 - no loading; 3 - Calu-1; 4 - H1975; 5 - H1650; 6- H1355; 7 - H1299; 8 - H1184; 9 - HCC827; 10 - H727; 11 - A549; 12 - H460; 13 - no loading; 14 – molecular weight marker for 140 kDa. This is one representative out of 3–5 independent experiments.

Article Snippet: Mouse anti-human CD22 MAbs (HB22-7 or RFB4) and chimeric anti-human CD20 MAb, RituximabTM (Biogen Idec, Inc., San Diego, CA), were added in a final volume of 200 μL at concentrations ranging from 5 to 50 μg/mL (3.33-33.3 × 10 −8 M).

Techniques: Incubation, Control, Labeling, SDS Page, Expressing, Molecular Weight, Marker

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Cell lines were cultured in quadruplicate in 96-well plates at 2 × 105 cells/mL in a volume of 100 µL complete medium. The following day, mouse anti-human CD22 MAbs (HB22.7 or RFB4) were added in a final volume of 200 µL at a final concentration ranging from 5 to 50 µg/mL (3.33 × 10−8 to 3.33 × 10−7 M). Both ITs [RFB4 (anti-CD22)-dgA and RFT5 (anti CD25)-dgA] were added in a final volume of 200 µL at a final molar concentration ranging from 1 × 10−10 to 1 × 10−13 M. Cells were incubated for 72 hours and cell viability was calculated by using CellTiter 96® AQueous One Solution [blank columns - no treatment, media only; horizontally hatched columns - HB22-7 at 3.33 × 10−7 M; checkered columns - RFB4 at 3.33 × 10−7 M; black columns - RFB4 (anti-CD22)-dgA at 1.0 × 10−11 M; light gray columns - RFT5 (anti-CD25)-dgA at 1.0 × 10−11 M]. The figure depicts the average ± SD of cell viability from three independent experiments.

Article Snippet: Mouse anti-human CD22 MAbs (HB22-7 or RFB4) and chimeric anti-human CD20 MAb, RituximabTM (Biogen Idec, Inc., San Diego, CA), were added in a final volume of 200 μL at concentrations ranging from 5 to 50 μg/mL (3.33-33.3 × 10 −8 M).

Techniques: Cell Culture, Concentration Assay, Incubation

Journal: Cancer research

Article Title: A Re-evaluation of CD22 Expression by Human Lung Cancer

doi: 10.1158/0008-5472.CAN-13-1436

Figure Lengend Snippet: Immunostaining performed on serial sections of lung squamous cell carcinoma shows pan cytokeratin 5/6/18 positive cancer cells (A) and CD22 positive lymphocytes (B). Immunostaining performed on serial sections of lung adenocarcinoma shows thyroid transcription factor-1 positive cancer cells (C) and CD22 positive lymphocytes (D). Scale bars = 40 µm.

Article Snippet: Mouse anti-human CD22 MAbs (HB22-7 or RFB4) and chimeric anti-human CD20 MAb, RituximabTM (Biogen Idec, Inc., San Diego, CA), were added in a final volume of 200 μL at concentrations ranging from 5 to 50 μg/mL (3.33-33.3 × 10 −8 M).

Techniques: Immunostaining