Proteins & Enzymes for Cell Therapy

MHC Complexes, CAR-T Targets, VLPs, Cytokines, and Laminin 521

Overview

Cell therapy includes immunocellular therapy and stem cell therapy. It utilizes cells from the patient or a donor, which, after being cultured, expanded, activated, or genetically edited ex vivo, are reinfused into the patient. This stimulates or enhances the body's immune function, thereby achieving a method of controlling diseases. Based on different mechanisms of action, typical types of cell therapy include CAR-T, TILs, TCR-T, CAR-NK, etc.

Figure 1. General Process of Immunocellular Therapy [1][2].

KACTUS Proteins for Cell Therapy Development

Supporting you from discovery through manufacturing.

Immunization Proteins for Drug Discovery

During the drug discovery phase of cell therapy, especially for reprogrammed cell therapeutic drugs, such as immune cells loaded with CAR (Chimeric Antigen Receptor), it is often necessary to obtain the correct scFv (Single-chain Variable Fragment) sequence through immunization with relevant target antigen proteins. This enables the drug product cells to accurately identify tumor cells expressing antigens. KACTUS offers a series of high-quality target antigen protein options for customers to choose from.

MHC Complexes

Intracellular antigen targets are a major direction in cell therapy. Peptide segments, resulted from the degradation of intracellular antigens, form complexes with MHC and are presented on the cell surface for TCR recognition. Targeting MHC peptide complexes can achieve the killing of tumor cells.

KACTUS peptide-MHC complexes cover a variety of popular targets, and include monomers, tetramers, fluorescently-labeled tetramers, and other forms, which can be used for immunization or screening research in the development of T-cell therapeutic drugs. KACTUS also offers custom MHC expression, custom TCR expression, and peptide-ready MHCs to build your own peptide-MHC in house.

Figure 2. Through ELISA and SPR experiments, we have verified that the monomer of HLA-A*02:01&B2M&AFP (FMNKFIYEI) complex can bind separately with Anti-AFP (HLA-A*02:01) Antibody and HLA-A*02:01&B2M&AFP TCR. The EC50 and affinity constant are 7.6 ng/mL and 0.923 μM, respectively.

Figure 3. (Left) Through ELISA validation, it has been confirmed that gp100 TCR & CD3 scfv fusion protein can bind to the monomer of HLA-A*02:01&B2M&GP100 (YLEPGPVTA) complex, with an EC50 of 11.8 ng/mL. (Right) In the SPR analysis, gp100 TCR & CD3 scfv fusion protein can bind to the tetramer of the HLA-A*02:01&B2M&GP100 (YLEPGPVTA) complex, with an affinity constant of 0.196 nM.

CAR-T Target Proteins

KACTUS' high-activity CAR-T target proteins can be applied during the early animal immunization stage for antibody discovery.

Figure 4. CAR-T target proteins, Human CD19 and Human CD7 proteins, can both bind well with the corresponding antibodies.

VLP SuperAntigens

Cell therapy targets include multiple transmembrane proteins (such as the GPCR family) and some targets for which it is difficult to obtain ideal antibody sequences through direct immunization. Our virus-like particle (VLP) display technology can maintain the correct conformation of multiple transmembrane proteins while enhancing the immunogenicity of the protein. In addition, the VLP framework can be designed specifically for your antigen with our custom VLP services.

Proteins for Cell Therapy Production & Analysis

Production preparation is a crucial step to obtain immunocellular drugs and is also key to the successful registration of cell drugs. KACTUS provides high-activity cytokines for cell culture and cell matrix proteins required for stem cell differentiation.

Laminin 521 for Stem Cell Culture

KACTUS offers recombinant Laminin 521 at research-grade, preclinical-grade, and cell therapy-grade. Laminin 521 is an important matrix adhesion protein for maintaining stem cells in culture.

Figure 5. The cell culture matrix protein Laminin 521 can effectively promote the growth of human iPSCs.

Figure 6. The cell culture matrix protein Laminin 521 can maintain the differentiation potential of stem cells.

Cytokines for Cell Culture

Cell culture is a necessary step for the preparation of cell-based therapies. Immunotherapy involving T cells, NK cells, and stem-cell derived immunocytes requires the preparation of stem cells such as ESCs, MSCs, and iPSCs. KACTUS offers a variety of cytokine products for cell culture.

Figure 7. (Left) We demonstrate via cell analysis that our Human IL-15 can promote the proliferation of MO7e cells, with an ED50 of 1-2 ng/mL. (Right) Through cellular level analysis, Human Wnt Surrogate can induce the activity of the Topflash reporter gene in HEK293 cells, with an ED50 of 5.2 ng/mL.

Figure 8. (Left) Via cellular-level analysis, we demonstrate that in the presence of Wnt Surrogate (5 ng/mL), Human R-spondin 1 Protein induced the activity of the Topflash reporter gene in HEK293 cells, with an ED50 between 1.0-10.0 ng/mL. (Right) Via cellular-level analysis, we validated that Human IL-7 Protein can promote the proliferation of 2E8 cells, with an ED50 value between 0.1-0.5 ng/mL.

Fluorescent or Biotinylated Proteins For Antibody Screening & Cell Therapy Analysis

Proteins labeled with fluorescence or biotin can be used for various detection processes in the production and quality inspection of therapeutic cells, such as antibody screening (such as scFv, etc.), CAR affinity research, CAR-T positivity rate detection, TCR screening or detection, and Blocking Assay, etc. KACTUS provides high-quality proteins labeled with different fluorescent markers like FITC, PE, as well as biotin-labeled proteins, to meet the needs of various analytical scenarios including cellular level analyses.

Figure 9. Through cellular-level flow cytometry detection, we demonstrated that FITC-Compatible Human CD19 protein can specifically bind with Anti-CD19-CAR T cells.

Figure 10. Through cellular-level flow cytometry detection, we demonstrated the fluorescently labeled HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQC) tetramer complex can bind well with HLA-A*02:01&B2M&NY-ESO-1 TCR cells.

GMP-Grade CRISPR Enzymes for Cellular Gene Editing

Allogeneic off-the-shelf cellular therapy products, such as UCAR-T, require the use of gene-editing technologies to knock out genes related to immune rejection reactions on donor T cells, to avoid inducing immune rejection reactions like GvHD and HvGR.

KACTUS offers GMP-Grade CRISPR Cas9 and GMP-Ready Cas12a for genetic modification of cells, including UCAR-T cell therapy. Regulatory support documentation is available. Moreover, our GMP-Grade Cas9 has been registered with the FDA Drug Master files (DMF #036578). Additionally, check out our CRISPR Cas9 ELISA kit to detect residual Cas9 nuclease in your cell product.

Figure 11. (Left) Cas9 Nuclease is used for gene knockout in 293T cells, Jurkat cells, and T cells, and its gene knockout efficiency in various cell types is comparable to that of leading suppliers. (Right) Cas9 Nuclease In Vitro DNA Cleavage Experiment. The in vitro cleavage efficiency of Cas9 nuclease is equivalent across all three batches.

Cas9 ELISA Kit

Quantitatively detect the residue of Cas9 nuclease in extracellular or intracellular spaces of gene-edited cell drugs before reinfusion into the human body. The detection range of our Cas9 ELISA kit is 0.25 ng/mL - 16 ng/mL, with a sensitivity that can reach 0.125 ng/mL.

Figure 12. Polynomial fitting of the Cas9 ELISA standard curve. The R² is 0.9997.

GMP-Grade MaxNuclease for DNA/RNA Degradation

In gene therapy, MaxNuclease principally serves to eliminate contaminants like host DNA and plasmid DNA present during the pharmaceutical production process. This GMP-grade nuclease is produced within a facility maintaining stringent GMP-Grade production standards, avoiding the use of materials derived from animals. The analytical methods applied have undergone systematic validation, and the final enzyme is subject to extensive quality control release testing to ensure it meets the requirements for use from research and development through to large-scale commercial production. Additionally, KACTUS has submitted MaxNuclease to the FDA Drug Master Files (DMF #036799).

Figure 13. Virus harvest solution was treated with 25U/mL and 50U/mL endonuclease at 37°C for 2 hours, respectively. Detection of plasmid DNA (pDNA) residue (left) and host cell DNA (HCD) (right) residue was analyzed. KACTUS MaxNuclease has higher degradation activity versus Competitor B demonstrated by lower pDNA and HCD residue for both 25U/mL and 50U/mL working concentrations.

MaxNuclease ELISA Kit

The quantification of residual MaxNuclease in gene therapy drugs is achievable with our associated ELISA kit, which provides a wide detection range of 46.88pg/mL to 3000pg/mL and sensitivity reaching up to 23.44pg/mL.

Figure 14. Example standard curve for MaxNuclease ELISA kit.

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References

[1] Fang KK, Lee JB, Zhang L. Adoptive Cell Therapy for T-Cell Malignancies. Cancers (Basel). 2022 Dec 23;15(1):94.

[2] Mangal JL, Handlos JL, Esrafili A, Inamdar S, Mcmillian S, Wankhede M, Gottardi R, Acharya AP. Engineering Metabolism of Chimeric Antigen Receptor (CAR) Cells for Developing Efficient Immunotherapies. Cancers (Basel). 2021 Mar 5;13(5):1123.

[3] Maalej KM, Merhi M, Inchakalody VP, Mestiri S, Alam M, Maccalli C, Cherif H, Uddin S, Steinhoff M, Marincola FM, Dermime S. CAR-cell therapy in the era of solid tumor treatment: current challenges and emerging therapeutic advances. Mol Cancer. 2023 Jan 30;22(1):20.

Proteins & Enzymes for Cell Therapy

MHC Complexes, CAR-T Targets, VLPs, Cytokines, and Laminin 521

Overview

KACTUS Proteins for Cell Therapy Development

Immunization Proteins for Drug Discovery

MHC Complexes

CAR-T Target Proteins

VLP SuperAntigens

Proteins for Cell Therapy Production & Analysis

Laminin 521 for Stem Cell Culture

Cytokines for Cell Culture

Fluorescent or Biotinylated Proteins For Antibody Screening & Cell Therapy Analysis

GMP-Grade CRISPR Enzymes for Cellular Gene Editing

GMP-Grade MaxNuclease for DNA/RNA Degradation

Browse Cell Therapy Products by Category

MHC Complexes

Soluble TCR Expression

CAR-T Target Proteins

VLP Antigens

Cytokines

Peptide-Ready MHCs

Gene Editing Enzymes

MaxNuclease, GMP

Laminin 521

Cell Therapy Proteins

References

Custom Protein Production

Bulk Inquiry

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