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Updated: Jun 16, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
Combination of adenovirus and cross-linked low molecular weight PEI improves efficiency of gene transduction
Jianfeng Han1, Dong Zhao, Zhirong Zhong
1Key Laboratory of Drug Targeting and Novel Drug Delivery Systems, Ministry of Education, West China School of Pharmacy, Sichuan University, Chengdu, Sichuan 610041, People's Republic of China.
Abstract:
Recombinant adenovirus (Ad)-mediated gene therapy is an exciting novel strategy in cancer treatment. However, poor infection efficiency with coxsackievirus and adenovirus receptor (CAR) down-regulated cancer cell lines is one of the major challenges for its practical and extensive application. As an alternative method of viral gene delivery, a non-viral carrier using cationic materials could compensate for the limitation of adenovirus. In our study, adenovectors were complexed with a new synthetic polymer PEI-DEG-bis-NPC (PDN) based on polyethylenimine (PEI), and then the properties of the vehicle were characterized by measurement of size distribution, zeta potential and transmission electron microscopy (TEM). Enhancement of gene transduction by Ad/PDN complexes was observed in both CAR-overexpressing cell lines (A549) and CAR-lacking cell lines (MDCK, CHO, LLC), as a result of facilitating binding and cell uptake of adenoviral particles by the cationic component. Ad/PDN complexes also promoted the inhibition of tumor growth in vivo and prolonged the survival time of tumor-bearing mice. These data suggest that a combination of viral and non-viral gene delivery methods may offer a new approach to successful cancer gene therapy.
Insights
This study introduces a novel polymer-adenovirus complex (Ad/PDN) to improve cancer gene therapy. The Ad/PDN complex enhances adenoviral gene delivery, even in cancer cells with low CAR expression, leading to better tumor inhibition and survival rates.
Area of Science:
- Biotechnology
- Gene Therapy
- Nanomedicine
Background:
- Recombinant adenovirus (Ad)-mediated gene therapy shows promise for cancer treatment.
- Poor infection efficiency in cancer cells with down-regulated coxsackievirus and adenovirus receptor (CAR) limits Ad-based therapy.
- Non-viral cationic carriers offer an alternative for overcoming Ad limitations.
Purpose of the Study:
- To develop and characterize a novel non-viral carrier, PEI-DEG-bis-NPC (PDN), for complexing adenoviral vectors.
- To evaluate the gene transduction efficiency of Ad/PDN complexes in CAR-expressing and CAR-lacking cancer cell lines.
- To assess the in vivo efficacy of Ad/PDN complexes in inhibiting tumor growth and improving survival.
Main Methods:
- Complexation of adenovectors with the synthetic polymer PDN.
- Characterization of Ad/PDN complexes using size distribution, zeta potential, and transmission electron microscopy (TEM).
- In vitro gene transduction assays in various cell lines (A549, MDCK, CHO, LLC) and in vivo tumor growth inhibition studies.
Main Results:
- Ad/PDN complexes were successfully formed and characterized.
- Enhanced gene transduction was observed in both CAR-overexpressing and CAR-lacking cell lines, attributed to improved viral binding and uptake.
- Ad/PDN complexes significantly inhibited tumor growth and prolonged survival in tumor-bearing mice.
Conclusions:
- The Ad/PDN complex effectively overcomes the CAR-dependency of adenoviral gene delivery.
- Combining viral and non-viral strategies offers a promising approach for enhanced cancer gene therapy.
- This novel vector system holds potential for broader application in cancer treatment.
