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Custom CAR-T Cell Development & Validation

$8,750.00

Item Cat No.: BCTCAR

Application: CAR-T cell development

Reactivity: Cell culture

BiCell Scientific’s CAR-T Cell Service provides customized pre-clinical CAR-T Cell generation and validation services including monoclonal antibody sequencing, chimeric antigen receptor cloning, lentivirus packaging, mouse or human T cell transduction, and in vitro testing of CAR-T cells.

Chimeric antigen receptor T cells (also known as CAR T cells) are T cells that have been genetically engineered to produce an artificial T cell receptor for use in immunotherapy.

Chimeric antigen receptors (CARs, also known as chimeric immunoreceptors, chimeric T cell receptors or artificial T cell receptors) are artificial receptor proteins that have been genetically engineered to give T cells the new ability to target a specific protein on tumor cells. The receptors are chimeric because they combine the antigen-binding domain of a monoclonal antibody and the T cell activating domain of CD3ζ protein into a single receptor.

BiCell Scientific’s CAR-T Cell Service provides customized pre-clinical CAR-T Cell generation and validation services including monoclonal antibody sequencing, chimeric antigen receptor cloning, lentivirus packaging, mouse or human T cell transduction, and in vitro testing of CAR-T cells.

  • DNA sequencing of heavy chain and light chain of monoclonal antibody (including BiCell Scientific’s antibody sequencing service, #BCABSE)
  • Molecular cloning of single-chain variable fragment (scFv) of heavy and light chains of antibody (including BiCell Scientific’s antibody cloning service, #BCABSE)
  • Molecular assembly of chimeric antigen receptor with scFv domain, CD28 transmembrane domain, CD3ζ cytoplasmic signaling domain, and CD28 costimulatory molecule and cloning into pLenti-CAR-T2A-GFP vector
  • Molecular packaging of assembled CAR protein into lentivirus (including BiCell Scientific’s lentivirus production service, #BCFTLV)
  • Harvesting and activation of mouse or human T cells (mouse T cells are harvested from mouse spleen and separated from B cells with nylon wool fiber column and activated with CD3/CD28 beads; human T cells are harvested from PBMC or leukopack (customer provided) and separated from B cells with MACS and activated with CD3/CD28 beads)
  • Transduction of mouse or human T cells with lentivirus (mouse or human T cells are transfected with lentivirus in the presence of IL-2 on retronectin-coated plates)
  • Validation of CAR expression in T cells with GFP or other fluorescent proteins
  • In vitro assay of the cytotoxic effect of CAR-T cells on tumor cells or cultured cells expressing antigen proteins (premade lentivirus encoding firefly luciferase is used to transduce tumor cells or other primary cells to introduce firefly luciferase activity)

Customers will receive the following deliverables at the end of each project:

  • Validated scFv cDNA sequence with guaranteed folding stability and binding affinity
  • Fully assembled CAR protein expression vector cloned into lentiviral backbone
  • Validated pre-packaged lentivirus of 1ml at titer of 1×109/ml pseudotyped with VSV-G coat protein and able to transduce mouse or human T cells at titer of 1×106/ml
  • Transduced mouse or human T cells (1×106 cells)
  • Guaranteed CAR protein expression in mouse or human T cells
  • Guaranteed cytotoxic effects of CAR-T cells on tumor cells or cultured cells expressing antigen proteins
  • Luciferase killing assay report (default method) or other types of cytotoxic assay of customer’s choice

Add-on options

  • Suicidal CAR-T cells expressing a concatenated Herpes simplex virus thymidine kinase SR39 protein, that when bound with Ganciclovir, kills CAR-T cells
  • Immortalized CAR-T cells expressing a concatenated human telomerase reverse transcriptase (TERT), that immortalizes CAR-T cells.

For Research Use Only. Not for use in clinical diagnostics or therapeutics.

  • DNA sequencing of heavy chain and light chain of monoclonal antibody – 1-2 weeks
  • Molecular cloning of chimeric antigen receptor – 2-3 weeks
  • Lentivirus packaging and production – 2-3 weeks
  • Mouse or human T cells harvesting, separation and activation – 2-3 weeks
  • Transduction of T cells and validation of CAR protein expression in transduced T cells – 3-4 weeks
  • In vitro cytotoxic testing of CAR-T cells on tumor cells or other types of target cells – 3-4 weeks

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Be the first to review “Custom CAR-T Cell Development & Validation”

"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

Contact us for questions or custom requests!