Rituximab (anti-CD20)

Catalog No.A2009 Batch: A200907

Technical Data

CAS No.	174722-31-7
Formulation	100 mM Pro-Ac, 20 mM Arg, pH 5.0
Isotype	Human IgG1
Source	CHO cells
Storage (From the date of receipt)	Store the undiluted solution at 4°C in the dark to avoid freeze-thaw cycles
Purity	99%
Protein concentration	5.8 mg/ml
Endotoxin Level	＜1EU/mg

Biological Activity

Description

Rituximab (anti-CD20) is a chimeric anti-CD20 mAb that binds the CD20 antigen on B cells with a binding affinity of 5 nM, MW: 143.86 KD.

Targets

CD20 ^[1]

In vitro

Anti-CD20 IgG1 antibody (rituximab), induces strong antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) effects on CD20+ Raji cells with the EC50 at 0.0365 nM and 1.5 nM, respectively.^[1]

In vivo

A number of in vivo tumor models suggest the anti-tumor activity of rituximab is dependent, at least in part, on complement^[2]. Rituximab can deplete B cells for several months and, as such, could represent an effective therapy for B cell-mediated autoimmune diseases. Rituximab is now widely used in onco-haematology and is currently in development in several autoimmune diseases^[3].

Protocol (Only for Reference)

Cell Assay:	For ADCC experiments, PBMCs were isolated from healthy human donor blood to serve as effector cells and were plated with target Raji cells at E:T ratio of 50:1 in the presence of indicated concentrations of KPL-404, anti-CD20, or isotype antibodies. Target Raji cells were labeled with calcein-AM for detection of cell lysis prior to experimental set up. Following incubation for 4 hours at 37°C in 5% CO₂, supernatants from the experiment were analyzed. For CDC experiments, human complement was plated with calcein-AM labeled target Raji cells in the presence of indicated concentrations of KPL-404, anti-CD20, or isotype antibodies. Following incubation for 4 hours at 37°C in 5% CO₂, supernatants from the experiment were analyzed.
Animal Study:	Objective: To study the mechanisms of follicular lymphoma’s resistance to rituximab Animal Models: Female CB17 severe combined immunodeficient(SCID) mice were injected (s.c.) with rituximab-resistant lymphoma cells (derived from a human FL) Formulation: -- Dosages: 10mg/kg, weekly Administration: i.p. Reference: https://www.ncbi.nlm.nih.gov/pubmed/19188155 Objective: To investigate the therapeutic effect of rituximab in combination with L19-IL2 on B-cell malignancies Animal Models: CB17/lcr severe combined mmunodeficiency (SCID) mice were injected (s.c.) Ramos or DoHH-2 lymphoma cells Formulation: saline Dosages: 200 μg Administration: i.v. Reference: https://www.ncbi.nlm.nih.gov/pubmed/19005180 Rituximab can apply to immunodeficient mice (eg: SCID mice), lymphoma cell lines and other related assays (Only for Reference)

Cell Assay:

For ADCC experiments, PBMCs were isolated from healthy human donor blood to serve as effector cells and were plated with target Raji cells at E:T ratio of 50:1 in the presence of indicated concentrations of KPL-404, anti-CD20, or isotype antibodies. Target Raji cells were labeled with calcein-AM for detection of cell lysis prior to experimental set up. Following incubation for 4 hours at 37°C in 5% CO₂, supernatants from the experiment were analyzed. For CDC experiments, human complement was plated with calcein-AM labeled target Raji cells in the presence of indicated concentrations of KPL-404, anti-CD20, or isotype antibodies. Following incubation for 4 hours at 37°C in 5% CO₂, supernatants from the experiment were analyzed.

Animal Study:

Objective: To study the mechanisms of follicular lymphoma’s resistance to rituximab
Animal Models: Female CB17 severe combined immunodeficient(SCID) mice were injected (s.c.) with rituximab-resistant lymphoma cells (derived from a human FL)
Formulation: --
Dosages: 10mg/kg, weekly
Administration: i.p.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/19188155

Objective: To investigate the therapeutic effect of rituximab in combination with L19-IL2 on B-cell malignancies
Animal Models: CB17/lcr severe combined mmunodeficiency (SCID) mice were injected (s.c.) Ramos or DoHH-2 lymphoma cells
Formulation: saline
Dosages: 200 μg
Administration: i.v.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/19005180

Rituximab can apply to immunodeficient mice (eg: SCID mice), lymphoma cell lines and other related assays (Only for Reference)

References	[1] Muralidharan S, et al. J Pharmacol Exp Ther. 2022 Apr;381(1):12-21. [2] George J. Weiner, et al. Semin Hematol. 2010, 47(2): 115-123. [3] Cartron G, et al. Crit Rev Oncol Hematol. 2007, 62(1):43-52.

References

Selleck's Rituximab (anti-CD20) has been cited by 16 publications

Distinct CD16a features on human NK cells observed by flow cytometry correlate with increased ADCC [ Sci Rep, 2024, 14(1):7938]	PubMed: 38575779
TLR9 agonism differentially impacts human NK cell-mediated direct killing and antibody-dependent cell-mediated cytotoxicity [ Sci Rep, 2024, 14(1):14595]	PubMed: 38918496
Downhill running does not alter blood C1q availability or complement-dependent cytotoxicity of therapeutic monoclonal antibodies against haematological cancer cell lines in vitro [ Sci Rep, 2024, 14(1):28239]	PubMed: 39548231
Noninvasive PET imaging of tumor PD-L1 expression with 64Cu-labeled Durvalumab [ Am J Nucl Med Mol Imaging, 2024, 14(1):31-40]	PubMed: 38500749
Noninvasive PET imaging of tumor PD-L1 expression with 64Cu-labeled Durvalumab [ Am J Nucl Med Mol Imaging, 2024, 14(1):31-40]	PubMed: 38500749
Daratumumab induces cell-mediated cytotoxicity of primary effusion lymphoma and is active against refractory disease [ Oncoimmunology, 2023, 12(1):2163784]	PubMed: 36632565
Daratumumab induces cell-mediated cytotoxicity of primary effusion lymphoma and is active against refractory disease [ Oncoimmunology, 2023, 12(1):2163784]	PubMed: 36632565
Development and implementation of natural killer cell simultaneous ADCC and direct killing assay [ Heliyon, 2023, 9(12):e22991]	PubMed: 38125417
Rheumatoid factor IgM autoantibodies control IgG homeostasis [ Front Immunol, 2022, 13:1016263]	PubMed: 36341420
Beta-Lapachone Attenuates BMSC-Mediated Neuroblastoma Malignant Transformation by Inhibiting Gal-3/Gal-3BP/IL6 Axis [ J Pharmacol Exp Ther, 2022, 381(1):12-21]	PubMed: 35078863

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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