Interest in B cells has recently increased in immunotherapies using Engineered B cells or Tumour-Infiltrating Lymphocytes (TILs). However, current methods for B cell expansion are inefficient, time consuming and can lead to insufficient B cell numbers and persistent T cell exhaustion. AdipoGen’s B-Xpander™ protein improves current protocols for B cell stimulation by mimicking the natural interaction between CD40L and CD40 molecules on B cells.
B-Xpander™ is a potent enhanced Multimeric human CD40L cytokine, for B cell activation, B cell expansion (proliferation) and cell therapy applications. In-house production using a proprietary protocol and a characterised and certified CHO cell line allows the supply of the most active Multimeric human CD40L proteins in GMP-like format.
B-Xpander™ Quality Features:
- Production in characterised and certified CHO cell line (for GMP-grade manufacturing)
- High Bioactivity tested by ELISA/Cell-based Assays
- Verified Purity & Homogeneity by SEC
- Low Endotoxin Levels
- Batch-to-Batch Consistency
- Animal-component free Production
- No cross-linking Reagents necessary
B-Xpander™ CHO Cell Line:
The production of the B-Xpander™ uses a characterised and certified proprietary CHO cell line which has been tested on Master Cell Bank Biosafety and GMP compliance.
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 multimerisation domain sequence from mouse Adiponectin allows the multimerisation (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.
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, AdipoGen’s B-Xpander™ and Multimeric CD40L proteins do not need an enhancer (antibody) to multimerise and to be active.
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 multimerisation and maximal activity.
- B-Xpander™ is an efficient B cell activation factor.
METHOD: PBL cells were incubated in 96-well plates (2×105 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 analysed 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™, AdipoGen’s 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 tumour-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 tumour 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 recognise tumour-specific antigens, or they can be expanded to enrich for tumour-infiltrating lymphocytes (TILs) that have natural tumour-specific reactivity. The patient’s tumour cells can then interact with the enlarged TIL cells to screen effector TIL cells that can kill tumour 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 tumour-specific antigen improve tumour-specific TILs.
B-Xpander™ and Multimeric human CD40L Grades for your experiments:
| Product Name | PID | Protein Construct |
|---|
B-Xpander™
| B-Xpander™ CD40L (human) (rec.) (Animal Free) | AG-40B-0010AF | • Produced using animal component-free medium in characterised and certified CHO cells • GMP-like production • Ex vivo B Cell Expansion for Cell Therapy |
| B-Xpander™ CD40L (human) (rec.) (GMP-Grade) | Custom Please Inquire | • GMP-Production outsourced to CRO • Ex vivo B Cell Expansion for Cell Therapy |
Other Research Grades
| CD40L (human) (multimeric) (rec.) | AG-40B-0010 | • Produced using standard serum/medium in CHO cells • Ex vivo B Cell Expansion for Research Use |
| CD40L (human) (multimeric) (rec.) (Certified Serum Grade) | AG-40B-0010CSG | • Produced using certified serum/medium in CHO cells for traceability • Ex vivo B Cell Expansion for Cell Therapy and Research Use |
Originally posted on: https://adipogen.com/b-xpander
Caltag Medsystems is the distributor of Adipogen products in the UK and Ireland. If you have any questions about these products, please contact us.