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Anti-EGFRvIII CAR-T Preclinical in vivo Assay

Target Background

EGFRvIII, also known as de2-7EGFR or ΔEGFR, is one of most common extracellular mutation variants of epidermal growth factor receptor (EGFR). EGFRvIII is a 145kDa cell surface protein that has a shorter extracellular domain and extra cysteine residue with intracellular domain remain unchanged compared to EGFR. Its cellular function is associated with PI3K/Akt/mTOR pathway, which usually involves in regulating anaerobic glycolysis, lipogenesis and tumor cell proliferation. Previous studies indicate EGFRvIII is rarely detected in normal cells but is overexpressed in human malignancies such as glioblastoma multiforme, ovarian, breast and non-small-cell lung cancers. This feature makes EGFRvIII an ideal target for immunotherapies that avoids potential damage on normal tissues.

Anti-EGFRvIII CAR-T Cell Therapy

Therapeutic potential of EGFRvIII in brain and other tumors has been supported by many studies. Recent Phase I clinical trials are already being performed to determine the safety and efficacy of CAR-T-EGFRvIII in the treatment of patients with EGFRvIII+ glioblastoma. Creative Biolabs fully understands that CAR-T therapy development targeting EGFRvIII is a time-consuming and costly process; our scientist group is highly experienced and equipped with state-of-the-art facilities. We provide a wide range of services to assist you and your team to accelerate this process.

Anti-EGFRvIII CAR-T Preclinical in vivo Assay

The epidermal growth factor receptor variant III (EGFRvIII): Where wild things are altered.
The FEBS Journal,2013. 280 (21), 5350-5370.

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

Creative Biolabs offers comprehensive services to evaluate anti-EGFRvIII CAR-T cell therapy in vivo. Our technical group is very experienced and efficient in data production. Experimental techniques can be customized at customers’ requests or purpose-orientated.
Xenograft model of glioblastoma
Five to six week-old female BALB nude mice are given subcutaneous injection of transfected U87MG cell line expressing EGFRvIII (or D-270MG, SMA560 cell) that is propagated and naturally expresses EGFRvIII in left or right flank. Tumor size is measured twice a week. Mice receive CAR-T cells by intravenous injection via tail vein. Creative Biolabs is also able to conduct intracerebral implantation of EGFRvIII+ cells. Cells are incubated into corpus striatum in right hemisphere of nude mice via a burr hole to bregma after mice are anesthetized and fixed in stereotactic apparatus. Tumor sizes of intracerebral implantation model are measured by in vivo imaging or in postmortem analysis.
Xenograft model of Head and Neck squamous carcinoma
Creative Biolabs provides head and neck squamous carcinoma (HNSCC) mouse model by subcutaneously injection of EGFRvIII expressing HNSCC cell UM-22B in SD nude mice and followed by intravenous injection of engineered CAR-T cells. Tumor sizes are measured by Vernier caliper after 24 days.

In vivo Assay Parameters and Techniques

Efficacy Tests
Tumor remission monitored by tumor volume recording or bioluminescence imaging and survival curve tracking.
Viability and Bio-distribution Studies
Durability and bio-distribution are evaluated by bioluminescence imaging, immunochemistry staining and real-time PCR
Toxicity Evaluation
Pilot tolerated evaluation: route of administration, dosage, MTD
Clinical observation: body weight, food consumption, behavior and pathological signs
Cytokine storm surveillance (fever, hypertension, prolonged cytopenia)
Postmortem analysis
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.

Creative Biolabs fully understands researches of adoptive T cell therapy targeting EGFRvIII are time-consuming and costly. With our highly experienced technical groups and state-of-the-art facilities, we would like to accelerate these studies by providing our top-quality in vivo tumor models and efficient analyzing services.


  1. Miao, et al. "EGFRvIII-specific chimeric antigen receptor T cells migrate to and kill tumor deposits infiltrating the brain parenchyma in an invasive xenograft model of glioblastoma." PLoS One 9.4 (2014): e94281.
  2. Sampson, et al. "EGFRvIII mCAR-modified T-cell therapy cures mice with established intracerebral glioma and generates host immunity against tumor-antigen loss." Clinical Cancer Research 20.4 (2014): 972-984.
  3. Morgan, et al. "Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma." Human gene therapy 23.10 (2012): 1043-1053.

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