Mixed Lymphocyte Reaction (MLR) Assay

Creative Biolabs provides excellent services for the analysis and characterization of Fab antibody functions by our advanced antibody functional analysis platform. Our professional scientists can help global customers discover potential immune checkpoint functions. Particularly, we are happy to introduce our cost-effective mixed lymphocyte reaction (MLR) assay.

Background

Mixed lymphocyte reaction (MLR) is a useful model used by pharmaceutical and biotech organizations to study alloresponsiveness to histocompatibility antigens. In short, it mixes populations of lymphocytes together and measures the reaction that occurs. Using the MLR, we can measure the disparity in major histocompatibility complex (MHC) antigens between individuals. Particularly, potential immunoactivity or immunosuppressive effects of your drugs can be determined using this assay.

Fig.1 Mixed Lymphocyte Reactions (MLR) with centrally or peripherally tolerant lymphocyte populations in graft recipients.¹

To date, a few types of MLR have been developed to meet scientists’ guidelines. Among these methods, two main types of MLR, namely one-way MLR and two-way MLR, have been widely used to help researchers characterize their drug candidates’ potential ability to mount anti-tumor T cell responses in the tumor microenvironment. One-way MLR is a cell proliferation assay where one population of lymphocytes (responder cells) is stimulated to proliferate by another population of lymphocytes (stimulator cells, which have been rendered non-proliferative). On the contrary, the two-way MLR differs from the one-way MLR in that both of the cell populations are able to proliferate.

MLR Assay Provided by Creative Biolabs

With abundant experience in this field, Creative Biolabs has established an advanced platform to help global researchers determine the effects of antibodies on the ability of T cells to be activated and proliferate. Depending on your wishes we can either offer one-way MLR or two-way MLR. Alternatively, the MLRs can be performed with various sources of antigen presenting cells (APCs), which include but are not limited to monocytes, immature and mature monocyte-derived dendritic cells (DCs), and plasmacytoid DCs. What’s more, our method will provide analysis and interpretation of the effects of your biologics or small molecules on multiple endpoints from multiple allogeneic donors.

Workflow of Our MLR Assay

We provide a comprehensive service process ranging from cell isolation to data visualization. Backed up by our competitive expert team, we have confidence in delivering standard project reports in a short turnaround (3-4 weeks).

Cells Types in MLR Assay

• T-cells, and T-cell Subsets (CD4+, CD8+)
• Dendritic Cells (DCs)
• Mesenchymal Stem Cells (MSCs)
• Peripheral Blood Mononuclear Cells (PBMCS)
• Endothelial Cells
• Cardiomyocytes
• Biomaterials (tendon, bone, graft tissue, skin, etc.)

Selective Readouts

• Cytokine secretion: Cytokine secretion detection by ELISA, RT-qPCR, and multiplex cytokine analysis technology.
• Surface markers: Cell surface markers (CD40, CD80, CD83, CD86, etc.) detection by flow cytometry.
• Cell proliferation: T cell proliferation by flow cytometry (CFSE) and 3H thymidine incorporation.

Case Study

This research assesses the impact of commercialized drug on MSCs' immunomodulatory activity. It involves real-time PCR for gene expression analysis of MSCs treated with commercialized drug, and secretome profiling. The immunomodulatory activity is measured through mixed lymphocyte reaction (MLR) and migration assays, highlighting the potential therapeutic value of MSCs in modulating immune responses.

Format: 96-well plate

Measure: 3H-thymidine incorporation

Fig.3 commercialized drug influences MSC-mediated suppression of allostimulated PBMC proliferation in MLR assays.²

Advantages of Our Services

• Multiple sources of APCs and modes of their activation/maturation for option
• Both standard and customized service to meet every client’s requirements
• Best after-sale service with cost-effective price
• Professional technical assistance staffs

Frequently Asked Question

1. Q: What is the maximum number of samples that can be tested in your MLR assay?

   A: The MLR assay can accommodate different capacities based on the experiment type. For large-scale MLR experiments, you can test between 11 and 16 samples simultaneously. Medium-scale MLR experiments allow for testing of 5 to 10 samples at a time. For small-scale MLR experiments, you can measure between 1 and 4 samples concurrently.

Creative Biolabs has been devoted to the field of antibody function studies for over a decade and has established a series of technology platforms to integrate various antibody function analysis services. With extensive experience, abundant industry knowledge, and advanced technical skills, our scientists are fully confident in accomplishing even the most challenging projects to make your project a success. For more information, please feel free to contact us.

References

1. Loewendorf, Andrea I., Marie Csete, and Alan Flake. "Immunological considerations in utero hematopoetic stem cell transplantation (IUHCT)." Frontiers in Pharmacology 5 (2015): 282.
   Distributed under Open Access License CC BY 4.0, without modification.
2. Zielniok, Katarzyna, et al. "Vadadustat, a HIF prolyl hydroxylase inhibitor, improves immunomodulatory properties of human mesenchymal stromal cells." Cells 9.11 (2020): 2396.
   Distributed under Open Access License CC BY 4.0, without modification.

For Research Use Only.