B-Xpander™ - Unleash the Power of B Cell Expansion

B-Xpander™ – The Multimeric human CD40L Structure

• B-Xpander™ / Multimeric human CD40L is a high activity fusion protein construct in which the human CD40L extracellular domain (aa 116-261) is artificially linked via the collagen domain of ACRP30.

• The ACRP30headless multimerization domain sequence from mouse Adiponectin allows the multimerization (10-12 x trimers) of the construct.

• This construct very effectively simulates the natural membrane-assisted aggregation of human CD40L in vivo.

• MultimericLigands developed by AdipoGen Life Sciences are ideal for in vitro & ex vivo applications.

FIGURE: Structure of the B-Xpander™ or Multimeric human CD40L.

B-Xpander™ – B Cell Expansion (Proliferation)

B-Xpander™ increases proliferation of human primary CD19+ B cells as efficiently as other grades of Multimeric human CD40L. Unlike most recombinant CD40L proteins on the market, our B-Xpander™ and Multimeric CD40L proteins do not need an enhancer (antibody) to multimerize and to be active.

FIGURE: Measurement of B cell proliferation.

METHOD: Activity of B-Xpander™ and 2 different research grades of Multimeric hCD40L is measured by proliferation of human primary CD19+ B cells. Cells (5 x 10⁴/well) are grown in RPMI medium, glutamine, 10% FBS, IL-4 (human) (10ng/ml), IL-21 (human) (10ng/ml) and different concentrations of B-Xpander™ CD40L (human) (rec.) (Animal-Free) (#AG-40B-0010AF), CD40L (human) (multimeric) (rec.) (CSG; Certified Serum Grade) (#AG-40B-0010CSG), CD40L (human) (multimeric) (rec.) (#AG-40B-0010), or a control protein. Cell proliferation is quantified after 4 days using PMS/MTS (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega). Cells expanded by 10-fold after 4 days to reach 5 x 10⁵/well.

B-Xpander™ – B Cell Activation

• The multimeric structure of B-Xpander™ mimics the biological activity of membrane-bound human CD40L.

• B-Xpander™ is the most potent fusion protein alternative to activate human CD40.

• Using B-Xpander™, no enhancer (antibody) is required for multimerization and maximal activity.

• B-Xpander™ is an efficient B cell activation factor.

FIGURE: B-Xpander™ (Prod. No. AG-40B-0010AF) induces B cell activation.

METHOD: PBL cells were incubated in 96-well plates (2x105 cells/well in 100μl RPMI supplemented with 10% FCS) for 24 hours at 37°C with the indicated concentration of B-Xpander™ CD40L (human) (rec.) (Animal Free) (#AG-40B-0010AF), CD40L (human) (multimeric) (rec.) (#AG-40B-0010) or a trimeric human CD40L as a control. Cells were washed with PBS and stained with 2μl each CD86-PE and CD19-FITC in 50μl FACS buffer (PBS, 5% fetal calf serum, 0.02% azide) for 20 min. at 4°C in the dark. After two washes in FACS buffer, samples were then analyzed by flow cytometry.

B-Xpander™ – The Power of B Cell Expansion in Adoptive Cell Therapy

Embrace the future of immunotherapy research with Multimeric human CD40L-powered B cell expansion. Backed by rigorous scientific research and development, B-Xpander™, our cutting-edge Multimeric human CD40L-based B cell expansion tool, represents a major improvement in immunotherapy. During Adoptive Cell Transfer (ACT) immunotherapy, activation and expansion of B cells present with T cells in tumor-infiltrating lymphocytes (TILs) by B-Xpander™ leads to increased proliferation and greater abilities of T cells to fight cancer cells (Arnaud, et al. 2022).

LIT: Sensitive identification of neoantigens and cognate TCRs in human solid tumors: M. Arnaud, et al.; Nat. Biotechnol. 40, 656 (2022)

Adoptive Cell Therapy (ACT)

During ACT, T cells are isolated from a patient's blood or tumor tissue, expanded and activated ex vivo, and then re-infused back into the patient to target and eliminate cancer cells. T cells can be engineered to express chimeric antigen receptors (CARs) or T cell receptors (TCRs) that recognize tumor-specific antigens, or they can be expanded to enrich for tumor-infiltrating lymphocytes (TILs) that have natural tumor-specific reactivity. The patient's tumor cells can then interact with the enlarged TIL cells to screen effector TIL cells that can kill tumor cells.

Recently, it has been observed that B cells are a new key player in the Adoptive Cell Therapy. Higher presence of B cells infiltrated with TILs are associated with better TIL expansion and activation. B cells stimulated and amplified with B-Xpander™ and loaded with tumor-specific antigen improve tumor-specific TILs.

FIGURE: ACT workflow for TIL expansion.

• Produced using animal component-free medium in characterized and certified CHO cells
• GMP-like production
• Ex vivo B Cell Expansion for Cell Therapy

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• GMP-Production outsourced to CRO
• Ex vivo B Cell Expansion for Cell Therapy

• Produced using standard serum/medium in CHO cells
• Ex vivo B Cell Expansion for Research Use

• Produced using certified serum/medium in CHO cells for traceability
• Ex vivo B Cell Expansion for Cell Therapy and Research Use