Non-clinical and clinical studies that evaluate pharmacokinetic (ADME) and toxicology characteristics are necessary to determine the safety and efficacy of cell and gene therapy products. Studies on gene vector and transgene biodistribution, gene expression, protein expression, viral shedding, vector integration, and other areas are crucial for cell and gene therapy preclinical and clinical research.
The team at Avance Biosciences™ has nearly three decades of experience providing services to support cell and gene therapy biodistribution studies, including qPCR, ddPCR, and NGS. Throughout the years, we have designed and validated numerous TaqMan assays and analyzed massive quantities of animal tissue samples.
We specialize in assay development, assay qualification or validation, and sample testing that complies with GLP and CGMP regulations.
| Testing Services | Descriptions |
| Non-Clinical Studies | |
| Biodistribution of DNA viral vectors or transgenes | The biodistribution of viral vectors or transgenes is an important part of pre-clinical pharmacokinetics and toxicity studies. To support biodistribution studies, Avance offers GLP and Non-GLP qPCR, ddPCR, and NGS testing services. Our team has extensive experience in qPCR biodistribution assay development, validation, and sample testing. |
| Biodistribution of RNA transgenes | The biodistribution of RNA transgenes is typically a requirement for RNA-based cell and gene therapy products. To determine RNA transgene biodistribution, we provide RT-qPCR method with RNA standard curves to test genes that are not naturally present in testing animals. |
| Biodistribution of therapeutic cells | Determining the biodistribution of therapeutic cells is essential for understanding the efficacy and safety of a cell and gene therapy product. The team at Avance Biosciences™ has much expertise in developing sensitive and specific qPCR, ddPCR, and NGS methods to quantify human cells in animal tissues. |
| Gene expression of gene of interest | We have nearly three decades of experience supporting gene expression studies using RT-qPCR and other methods such as RNA-Seq. We provide both the ΔΔCt method and the RNA standard curve method for gene expression studies. |
| Protein expression of transgenes | Quantifying the protein expression of transgene products may be required if a cell or gene therapy transgene is detected in unintended tissues or organs. The experienced protein team at Avance can assist you in developing specific and sensitive ELISA and Western blot assays for measuring protein expression levels. |
| Shedding/excretion analysis | Determining assay specificity and sensitivity can be challenging in viral or bacterial shedding studies of animal urine, feces, and bodily fluids. Avance has developed effective extraction methods to ensure nucleoids have high recovery and minimal PCR inhibition. |
| Vector integration study | When high levels of vector or transgene are persistently detected in a biodistribution study, a vector gene integration study is warranted. Avance has developed a proprietary NGS-targeted sequencing method that can detect less than 50 copies of integrated events in 1 µg of host genomic DNA. |
| Gene editing site profiling | Avance offers quantitative NGS amplicon sequencing to support distribution and stability studies of therapeutic cells with edited genes. We have successfully designed and validated multiple NGS amplicon assays and rhAmpSeq assay panels to support non-clinical studies under GLP regulations. |
| Viral vector titer determination for dosing | To accurately titer viral vector titers at various doses, Avance has developed accurate and robust assays that utilize a proprietary viral-handling process. Our procedures reduce viral aggregation and viral adhesion to containers and pipet tips, maximizing reproducibility and recovery. |
| Clinical Studies | |
| Transgene RNA expression study | Assessing transgene expression regulation is of great importance for evaluating cell and gene therapy safety and efficacy. The Avance team is experienced in analyzing patient blood samples for RNA expression using GLP-compliant qPCR and ddPCR methods. |
| Gene editing site profiling | Avance offers quantitative NGS amplicon sequencing that evaluates the persistence and stability of gene editing sites during human trials. We have successfully validated multiple GLP-compliant NGS amplicon assays to support clinical trials. |
| Shedding/excretion analysis | The Avance team is experienced in developing and validating qPCR, ddPCR, and NGS amplicon assays with efficient extraction methods that ensure high and consistent recovery of various cell and gene therapy products in human urine, stool, nasal swab, throat swab, and many other sample types. |
| Immunogenicity study | Treatment with cell and gene therapy products may result in immune responses such as elevated cytokine levels, antibodies targeting the drug product, and neutralizing antibodies. Contact us to discuss how our experienced scientific team can assist you with measuring the immunogenicity of your cell or gene therapy product. |
The team at Avance Biosciences™ has nearly three decades of experience providing services to support biodistribution studies for cell and gene therapies, including qPCR, ddPCR, and NGS. Throughout the years, we have designed and validated numerous TaqMan assays and analyzed massive quantities of animal tissue samples.
Assay Design and Validation
We follow FDA guidance and industry standards for PCR-based assays to measure the biodistribution of gene therapy.
- GLP for biodistribution as part of toxicity studies and non-GLP for stand-alone biodistribution studies.
- LOQ is ≥ 50 copies/µg of host genomic DNA with 95% confidence.
- A third well spike is included to assess potential PCR inhibition for every test sample .
- PCR efficiency is between 90 and 100%, with an R2 > 0.99.
- Assays are qualified and/or validated following ICH guidelines and FDA guidance.
- Stringent process controls are used to prevent cross-contamination.
Experience with various vectors, animals, and tissues
Whether you are working on plasmid-based or AAV gene therapies, CAR-T cell therapeutics, or iPSC-based therapies, Avance Biosciences™’ experienced team of scientists has seen it all. We can assist you in developing specific and sensitive assays for your biodistribution studies.
| Adeno associated virus (AAV) | CAR-T |
| Adenovirus (Ad) | TCR |
| Poxvirus | CAR-NK |
| Vaccinia virus | iPSC |
| Poliovirus | LNP encapsulated sgRNA/RNA |
| Lipid nanoparticles | Plasmid |
| Salmonella | Exosome |
Experience with various animals and tissue types
Our team is experienced with all common animal models used in biodistribution studies, including mouse, rat, monkey, rabbit, pig, and canine. In addition to the standard biodistribution study tissue panels (blood, bone marrow, injection site(s), testes, ovaries, brain, liver, kidneys, lung, heart, muscle, spleen and many others), we have experience working with almost every major organ as well as different sections of target and non-targeted organs and tissues. We routinely receive and test thousands of animal tissue and blood samples every month for biodistribution, gene expression, and protein expression studies!
| Adrenals | Epididymis | Gall Bladder | Mammary gland | Prostate | Spleen | Tongue |
| Aorta | Esophagus | Galt | Meninges | Prostate Gland | Sternum marrow | Trachea |
| Blood | Eyes | Heart | Muscle | Rectus femoris | Stomach | Trigeminal Ganglia |
| Bone Marrow | Feces | Kidney | Nerve | Salivary gland | Tail | Tumor |
| Brain | Gall Bladder | Larynx | Ovary | Seminal vesicles | Testes | Ureter |
| Cervix | Galt | Liver | Oviduct | Serum | Throat | Urinary Bladder |
| CSF | Heart | Lung | Pancreas | Skin | Thymus | Urine |
| DRG | Intestine | Lymph Node | Pituitary | Spinal cord | Thyroid | Uterus |
Extraction Recovery
To effectively evaluate cell and gene therapy vector distribution and persistence, the methods used for vector or transgene DNA or RNA extraction must be of high yield and purity. Although evaluating vector or transgene DNA or RNA extraction recovery is not explicitly stated in regulatory guidance documents, regulatory agencies do require the evaluation of analyte recoveries in both method development and validation.
Most extraction methods and kits are capable of extracting vector and host nucleic acids with high efficiency and good quality. However, certain viral vectors, such as poxvirus, are notoriously difficult to lyse, leading to low transgene recovery. Additionally, most commercial blood extraction methods suggest a short lysis period (less than 30 minutes), exacerbating the problem of incomplete vector lysis.
Avance Biosciences™’ scientists have creatively resolved several technical issues associated with vector transgene recovery methods by implementing:
- Routine use of stronger lysis conditions and an extended lysis process to ensure high recovery of viral vector DNA and RNA.
- Proprietary animal and human blood extraction methods extend lysis without coagulating the blood.
Our Customers Say…
Avance Biosciences™ (AB) has delivered the Sequencing data in a record time. Very impressive! We have benefited from AB’s hard work and are very grateful for that. AB team should feel proud of themselves. We thank AB team for their support and contributions, much appreciated.
Head of Quality Control, Microbial Manufacturing Service, California
Your team met the timelines we needed and did exceptional technical work. I will definitely make my management aware that Avance is a very capable partner for our bioanalytical needs.
Bioanalytical Senior Project Manager, California
My experience with Avance has been nothing but positive… I will keep sending work your way as I appreciate the attention your team provides and Avance’s creative technical team. See you soon.
Director Individualized Medicines, California
Avance has been the ideal partner in helping Mission Bio and our client,… be the first in the world to GMP qualify our single cell % transduction assay for a lentiviral gene therapy product. Excellent execution on the part of the Avance team. We look forward to more collaborations in the future.
Senior Director, Cell & Gene Therapy Business, California
From the preliminary conclusion of this work, we are already pleased to thank you for your customer-oriented approach and proactive communication in the course of this study. We are happy to see the profound difference in business mindset between your organization and our previous vendor…
Project Manager, Belgium
Thank you so much for successfully completing this important project for us on time. We are very satisfied with the final report and thank you for taking the extra effort to customize the report format. We will definitely use your lab for future projects.
Program Manager, California
It has been a pleasure working with you and your team. The project was carried out to the highest standard and delivered in a timely manner. I will definitely recommend Avance Biosciences™’ services to colleagues and friends.
Senior Scientist, Singapore
When you partner with Avance Biosciences™, you gain a CRO partner that is creative, collaborative and dedicated to sound science with a focus on the regulatory requirements our partners require. Contact our technical staff to discuss how we can support you in your project!
