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BD Horizon™ BV605 Biosimilar Anti-Human CD20
Clone Obinutuzumab297.rMAb (also known as Obinutuzumab N297A Biosimilar.rMAb) (RUO)

Two-color flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415) and with either BD OptiBuild™ BV605 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756233; Left Plot) or BD Horizon™ BV605 Biosimilar Anti-Human CD20 (Obinutuzumab Biosimilar) antibody (Cat. No. 570972/571043; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ Software.

Multiparameter flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood leukocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD OptiBuild™ BV605 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756233; Left Plot) or BD Horizon™ BV605 Biosimilar Anti-Human CD20 (Obinutuzumab Biosimilar) antibody (Cat. No. 570972/571043; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ Software.



Two-color flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415) and with either BD OptiBuild™ BV605 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756233; Left Plot) or BD Horizon™ BV605 Biosimilar Anti-Human CD20 (Obinutuzumab Biosimilar) antibody (Cat. No. 570972/571043; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood leukocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD OptiBuild™ BV605 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756233; Left Plot) or BD Horizon™ BV605 Biosimilar Anti-Human CD20 (Obinutuzumab Biosimilar) antibody (Cat. No. 570972/571043; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ Software.

Two-color flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415) and with either BD OptiBuild™ BV605 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756233; Left Plot) or BD Horizon™ BV605 Biosimilar Anti-Human CD20 (Obinutuzumab Biosimilar) antibody (Cat. No. 570972/571043; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ Software.

Multiparameter flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood leukocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD OptiBuild™ BV605 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756233; Left Plot) or BD Horizon™ BV605 Biosimilar Anti-Human CD20 (Obinutuzumab Biosimilar) antibody (Cat. No. 570972/571043; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed using a BD Fortessa™ X-20 Flow Cytometer System and FlowJo™ Software.


BD Horizon™ BV605 Biosimilar Anti-Human CD20

BD Horizon™ BV605 Biosimilar Anti-Human CD20

Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- For U.S. patents that may apply, see bd.com/patents.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Clones without a listed workshop have not been investigated in an HLDA workshop to receive a CD nomenclature. We use “CD” provisionally when our internal testing indicates that this clone binds to the same CD antigen as workshopped clones.
The Obinutuzumab297.rMAb (also known as, Obinutuzumab N297A Biosimilar.rMAb) is a research grade humanized recombinant human IgG1, kappa antibody that specifically recognizes the extracellular domain of human CD20 similarly to the therapeutic Obinutuzumab antibody. The Obinutuzumab297.rMAb uses the same variable region sequences as Obinutuzumab, combined with constant region sequences derived from consensus sequences of human IgG1, kappa. The asparagine at position 297 of the constant heavy chain has been replaced with alanine (N297A) to further reduce Fc receptor interactions of this reagent. Obinutuzumab triggers target cell death through multiple mechanisms including complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and apoptosis. The therapeutic Obinutuzumab has been approved to treat human B cell malignancies, autoimmune disorders, and transplant rejection. CD20 is a 33-37 kDa, unglycosylated four-transmembrane phosphoprotein with a cytoplasmic N-terminus and C-terminus that is encoded by MS4A1 (Membrane-spanning 4-domains, subfamily A, member 1). CD20 is expressed on pre-B-cells, naive and activated B cells, memory B cells, and at lower levels on a small subset of T cells but is absent from plasmablasts and plasma cells. It is also expressed on most malignant B cells and CD20-positive T-cell lymphomas. CD20 functions in mediating calcium transport and B cell activation, differentiation, and survival.
The Obinutuzumab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

Development References (5)
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Almasri NM, Duque RE, Iturraspe J, Everett E, Braylan RC. Reduced expression of CD20 antigen as a characteristic marker for chronic lymphocytic leukemia. Am J Hematol. 1992; 40:259-263. (Biology).
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Brekke OH, Sandlie I. Therapeutic antibodies for human diseases at the dawn of the twenty-first century.. Nat Rev Drug Discov. 2003; 2(1):52-62. (Biology). View Reference
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Hultin LE, Hausner MA, Hultin PM, Giorgi JV. CD20 (pan-B cell) antigen is expressed at a low level on asubpopulation of human T lymphocytes. Cytometry. 1993; 14:196-204. (Biology).
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Tobinai K, Klein C, Oya N, Fingerle-Rowson G. A Review of Obinutuzumab (GA101), a Novel Type II Anti-CD20 Monoclonal Antibody, for the Treatment of Patients with B-Cell Malignancies.. Adv Ther. 2017; 34(2):324-356. (Biology). View Reference
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Zola H, Swart B, Nicholson I, Voss E. CD20. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:72.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.