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Anti-MUC1 CAR-T Preclinical in vivo Assay

Target Background

Mucin (MUC1) is a glycoprotein with extensive O-linked glycosylation of its extracellular domain and expressed on the apical surface of epithelial cells in organs such as lung, stomach, intestine and eye. Under normal physiological conditions, mucins prevent pathogen invasion by oligosaccharides in the extracellular domain, which plays an important role in keeping pathogens from reaching the cell surface. While in the cancerous scenario, several changes occur including transcriptional upregulation, lack of polarity and dysregulation of O-linked glycosylation. Correspondingly, those changes result in the overexpression of MUC1, easier access to the immune system as well as a preponderance of shorter glycan adjunctions (Thomsen-nouveau (Tn), sialyl Tn (STn), Thomsen-Friedenreich (T), and sialyl-T (ST)) in the breast, ovarian, lung and pancreatic cancer cells, which render MUC1 a potential “holy grail” among immunotherapeutic targets.

Anti-MUC1 CAR-T Cell Therapy

Multiple clinical trials are being conducted for anti-MUC1 engineered CAR-T cells, which employ different CAR design from utilization of an SM3 scFv, P28z CAR, to the recent 5E5 scFv. Delayed tumor progression has been shown in preclinical in vivo study of breast cancer, T-cell leukemia and pancreatic cancer. In Phase I clinical study of MUC1-positive metastatic seminal vesicle cancer, promising results were also observed, which constitutes a proof of concept that aberrant tumor glycosylation could be specifically targeted by CAR-T cell therapy to control both hematological and solid malignancies. Of note, much remains to be elucidated in terms of signaling potency, mechanisms of action, trafficking pattern and safety profiles. Combination therapy strategy with immune checkpoint blockers, specific chemokines, and vessel-normalizing agents are crucial aspects need to be studied as well.

Anti-MUC1 CAR-T Preclinical in vivo Assay

Selective Recognition of the Tumor-Associated Hypoglycosylated Forms Tn-STn on MUC-1 by T Cells Engineered to Express the Chimeric Antigen Receptor 5E5
Immunity. 2016 Jun 21;44(6):1248-50.

Animal Models for in vivo Study of anti-MUC1 CAR-T Cell Therapy

Based on the full-fledged CAR-T in vivo study strategies at Creative Biolabs, we offer the most exquisite animal models with prominent expression of MUC1 including but not limited to the following categories.
T cell leukemia xenograft models
Systematic T cell leukemia models at Creative Biolabs are established by implantation of luciferase-labeled Jurkat or K562 cells (Tn-MUC1 positive) into NSG mice via tail-vein injection. Anti-MUC1 CAR-T cell therapy is initiated when the mean total flux of the tumor reaches between 107 and 108 photons/sec. Disease progression and therapeutic effects are monitored regularly through bioluminescence imaging.
Pancreatic cancer/breast cancer/ovarian cancer xenograft models
Pancreatic cancer models at Creative Biolabs are established by intraperitoneal injection of luciferase-labeled human Capan-2 or Hs766T cells (Tn-MUC1 positive) into NSG mice. Disseminated breast cancer models implanted with either luciferase-labeled MDA-MB-453 or MCF-7 cells (Tn-MUC1 positive) are also available at Creative Biolabs. Tumor cells are inoculated intraperitoneally into NSG mice. When the mean total flux of the tumor reaches between 107 and 108 photons/sec, anti-MUC1 CAR-T cells as well as corresponding control T cells are injected into the lateral tail vein. Disease progression and therapeutic effects are monitored regularly through bioluminescence imaging. Due to the fact that there exists positive expression of Tn-MUC1 in some primary ovarian cancer, Creative Biolabs is willing to assist our customers in establishing primary ovarian cancer models as well as related in vivo assays.

In vivo Assay Parameters and Techniques

At Creative Biolabs, we offer the most exquisite and comprehensive service platform for anti-MUC1 CAR-T cell therapy research.
Efficacy Test
Tumor remission monitored by tumor volume recording or bioluminescence imaging and survival curve tracking.
Viability and Bio-distribution Studies
Durability, GLP-compliant bio-distribution studies
Toxicity Evaluation
Pilot tolerability (MTD, The route of administration, Dose regimen/response/onset)
Clinical observation (body weight, feed consumption, ophthalmologic and clinical pathology)
Cytokine storm surveillance (fever, hypertension, prolonged cytopenia)
Complete necropsy, organ weight
Tumorigenicity study
GLP-Compliant Preclinical Test
All our experiments are performed by well-trained and experienced technicians in a GLP-compliant and IACUC-regulated facility.

At Creative Biolabs, we are dedicated to establish the most reliable tumor models and the most accurate research procedures by keeping up with the science pace towards CAR-T in vivo study in order to promote the transformation of preclinical researches to clinical therapies.


  1. 1. Agnusdei, V., et al. "Therapeutic antibody targeting of Notch1 in T-acute lymphoblastic leukemia xenografts." Leukemia 28.2 (2014): 278-288.
  2. 2. Posey, et al. "Engineered CAR-T Cells targeting the cancer-associated Tn-glycoform of the membrane mucin MUC1 control adenocarcinoma." Immunity 44.6 (2016): 1444-1454.
  3. 3. Wilkie, et al. "Retargeting of human T cells to tumor-associated MUC1: the evolution of a chimeric antigen receptor." The Journal of Immunology 180.7 (2008): 4901-4909.
  4. 4. Blidner, et al. Rabinovich. "Driving CARs into Sweet Roads: Targeting Glycosylated Antigens in Cancer." Immunity44.6 (2016): 1248-1250.
  5. 5. You, et al. "Phase 1 clinical trial demonstrated that MUC1 positive metastatic seminal vesicle cancer can be effectively eradicated by modified Anti-MUC1 chimeric antigen receptor transduced T cells." Science China Life Sciences 59.4 (2016): 386-397.
  6. 6. Maher, et al. "CAR mechanics: driving T cells into the MUC of cancer." Cancer research 69.11 (2009): 4559-4562.

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