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Adenovirus Service

Q: Which cells can adenovirus infect?

Adenovirus has broad-spectrum infection capabilities, including most mammalian cell lines (such as HEK293, HeLa, A549, U87, etc.), though infection efficiency is relatively lower for stem cells or certain primary cells.

Q: What is the relationship between adenovirus titer and infection efficiency?

Higher titers can improve infection efficiency, but appropriate MOI (Multiplicity of Infection) selection should be considered to avoid excessive cytotoxicity.

Q: How should adenovirus be stored?

Long-term preservation at -80°C is recommended, avoiding repeated freeze-thaw cycles; short-term storage at 4°C is acceptable before use (within 1-2 weeks), but not recommended beyond this timeframe.

Q: Can you provide GMP-grade adenovirus?

Currently, we provide research-grade adenovirus services. For GMP production (for clinical trials or IND submissions), please contact us for customized evaluation.

Q: What are the differences between AdMax and AdEasy systems?

Comparison Item AdMax System AdEasy System Shuttle Plasmid pHBAd series pHBAd series Packaging System AdMax AdEasy Backbone Plasmid pBHGlox(ΔE1,3)Cre pAdEasy-1 Packaging Cells 293A cells 293A cells Packaging Strategy Direct transfection into 293A cells after linearization Recombination in E. coli after linearization, followed by transfection Linearization Required for Packaging No, vector is expressed directly in 293A cells Yes, linearization after recombination, then transfection into 293A cells Titer Detection Plaque assay Plaque assay Purification Method CsCl density gradient centrifugation CsCl density gradient centrifugation Advantages Simple operation, high virus output rate, short cycle High packaging success rate, low recombination false positives Disadvantages Risk of recombination false positives Complex packaging process, longer cycle

Adenovirus OverviewService PortfolioTechnical AdvantagesFAQ

Creative Biogene specializes in providing high-quality adenovirus vector services, encompassing the construction, amplification, and purification of replication-deficient adenoviruses and helper-dependent adenoviruses (HDAd), supporting various serotypes including Ad5 and Ad5/F35. Leveraging our advanced QVirus™ platform, we have established mature and stable methodologies for vector construction, in vitro recombination, cellular packaging, recombinant adenovirus amplification, and gradient ultracentrifugation purification, complemented by systematic quality control and identification processes to ensure high purity and high-titer viral products.

Introduction to Adenovirus

Adenovirus (Ad) is a non-enveloped double-stranded DNA virus with a genome length ranging from 26 to 46 kilobase pairs (kb) and an icosahedral nucleocapsid structure. Since the 1980s, adenoviruses have become essential tools in gene therapy, vaccine development, and cancer treatment due to their efficient gene delivery capabilities, broad host range, and excellent genetic stability.

Several adenovirus vector-based vaccines and therapeutic products have received global regulatory approval, such as the COVID-19 vaccine (AstraZeneca's Vaxzevria) and Ebola virus vaccine (Janssen's Ad26.ZEBOV), validating the clinical value of adenovirus technology.

Classification of Adenovirus Vector Systems

Based on the number of viral genes retained and dependency levels, adenovirus vector systems are generally classified into three categories:

Replication-competent Adenovirus

Natural viruses without deletion of key replication genes or viruses designed for specific cell lysis
Primarily used for oncolytic virus research and other specialized applications
Not used for conventional gene delivery

Replication-deficient Adenovirus

Deletions in E1 and/or E3 regions, currently the most commonly used adenovirus vector type
Features excellent infection capability and approximately 7.5 kb foreign gene carrying capacity
Suitable for cellular and animal model experiments

Helper-dependent Adenovirus

Almost all viral coding sequences removed, retaining only ITRs and ψ signal
Requires helper virus for packaging
Higher safety profile, lower immunogenicity, and transgene capacity up to 37kb

Adenovirus Service Portfolio

With the continuous development of gene therapy, biopharmaceuticals, and basic scientific research, adenovirus has become one of the most commonly used viral vectors due to its efficient transfection capability, non-integration into the host genome, and relatively large vector capacity. Creative Biogene, with years of viral vector research and production experience, has established an advanced adenovirus vector platform, providing customized adenovirus vector services covering basic research, animal experiments, preclinical studies, and commercial applications.

Service Offerings

Fiber-Modified Ad5-RGD Adenovirus Production

The Ad5-RGD adenovirus features RGD peptide insertions in the fiber protein to enhance infection efficiency, especially in tumor-targeting applications.

Fiber-Modified Ad5/35 Adenovirus Production

The Ad5/35 chimeric adenovirus combines an Ad5 backbone with Ad35 fiber to improve the transduction of hard-to-infect cells, such as hematopoietic cells.

Helper-Dependent Adenovirus (HDAd) Production

Helper-dependent adenoviruses (HDAd) remove all viral coding regions, offering up to 36 kb of transgene capacity with low immunogenicity for long-term expression.

Adenovirus Production & Purification

We provide a full adenovirus production and purification service, delivering high-titer and high-purity virus with excellent transfection performance.

Adenovirus Amplification & Purification

Our amplification and purification service supports scalable production of adenoviral vectors for both in vitro and in vivo studies.

Adenovirus Titration

Adenovirus titration is conducted using TCID50, qPCR, and immunofluorescence to quantify viral titers for diverse research needs precisely.

Technical Platform

Creative Biogene has established a comprehensive adenovirus vector construction and production platform, with core modules including:

1. Vector Construction System

Gene cloning and vector construction (Gateway, Gibson, and other technologies)
Multiple cistron or IRES/2A peptide expression strategies
Optional tags: Flag, HA, His, GFP, mCherry, etc.

2. Packaging Cell Lines and Virus Production

HEK293/293A cell lines for E1/E3-deleted Ad packaging
PER.C6 cells supporting large-scale production and clinical-grade applications
Serum-free cultivation system
3D suspension culture and microcarrier amplification processes supporting scale-up production

3. Purification and Concentration Technologies

CsCl ultracentrifugation (research grade)
Chromatographic purification (preclinical grade): anion exchange, density gradient chromatography
UF/DF ultrafiltration concentration systems

4. Technical Optimization and Innovation

Enhanced Safety

Minimized replication risk: Genomic engineering significantly reduces homologous recombination probability, meeting stringent regulatory standards
Serum-free cultivation: Eliminates the risk of exogenous pathogen introduction from animal components

Production Efficiency Optimization

High-density cell cultivation: Suspension culture technology supports 15-20×10⁶ cells/mL, increasing virus yield by 3-5 fold compared to adherent systems
Rapid process scale-up: Modular reactors and single-use technology shorten the process development cycle to 6-8 weeks
Single-use consumables: Reduces cleaning validation and equipment maintenance costs, suitable for parallel production of multiple product lines

Cost-Benefit Analysis

High recovery rate: Overall process recovery >40% (compared to approximately 30% for traditional processes), reducing material and time costs

Deliverables

Shuttle plasmid containing the target gene
Recombinant adenovirus plasmid
Detailed experimental report (including specific experimental procedures, experimental images, etc.)
Integrated vector sequencing report for IND application (optional)

Service Workflow

Creative Biogene provides one-stop adenovirus services from vector design to purification delivery, with the standard process as follows:

Project Evaluation and Design

Client provides target gene sequence
We assist with design optimization

Gene Synthesis and Vector Construction

High-fidelity synthesis and cloning methods
Ensuring sequence accuracy

Adenovirus Packaging and Amplification

Transfection and amplification in 293-series cells

Virus Purification and Testing

Provision of high-purity virus
Comprehensive quality testing

Virus Delivery and After-Sales Support

Lyophilized or liquid virus storage options available
Recovery service support

Quality Control System

Creative Biogene has established a comprehensive virus quality control process, covering the following key indicators:

Test Item	Method	Acceptance Criteria
Titer Determination	qPCR/ELISA/TCID50	Customer-specified range
Transgene Expression	Western blot / qPCR / IF	Clear expression
Purity	SDS-PAGE / HPLC	≥95%
Endotoxin Level	LAL test	<5 EU/mL
Sterility	Culture method	Sterile
Mycoplasma Detection	qPCR or culture method	Negative
Host Residual DNA/Protein	PicoGreen / ELISA	Compliant with standards

Technical Advantages

Highly Customizable

Supports any sequence, multi-gene construction, reporter gene co-expression

High-Titer Virus

Routinely reaches titers ranging from 10⁹ to 10¹² vp/mL

Stringent Quality Control Standards

Covers multiple indicators including transgene expression, titer, purity, endotoxin, sterility, etc.

GMP-Compatible Platform

Meets requirements for preclinical research and IND submissions

Multiple Modification Options

RGD, fiber replacement, targeting modifications supporting personalized therapeutic research

Extensive Project Experience

Serviced hundreds of virus development projects, covering cancer, cardiovascular, neurological, metabolic, and other disease areas

Contact Us

If you have any questions or requirements regarding our adenovirus services, please feel free to contact our technical support team. Our professional team will provide customized adenovirus solutions to meet your research and development needs.

FAQ

Which cells can adenovirus infect?

Adenovirus has broad-spectrum infection capabilities, including most mammalian cell lines (such as HEK293, HeLa, A549, U87, etc.), though infection efficiency is relatively lower for stem cells or certain primary cells.

What is the relationship between adenovirus titer and infection efficiency?

Higher titers can improve infection efficiency, but appropriate MOI (Multiplicity of Infection) selection should be considered to avoid excessive cytotoxicity.

How should adenovirus be stored?

Long-term preservation at -80°C is recommended, avoiding repeated freeze-thaw cycles; short-term storage at 4°C is acceptable before use (within 1-2 weeks), but not recommended beyond this timeframe.

Can you provide GMP-grade adenovirus?

Currently, we provide research-grade adenovirus services. For GMP production (for clinical trials or IND submissions), please contact us for customized evaluation.

What are the differences between AdMax and AdEasy systems?

Comparison Item	AdMax System	AdEasy System
Shuttle Plasmid	pHBAd series	pHBAd series
Packaging System	AdMax	AdEasy
Backbone Plasmid	pBHGlox(ΔE1,3)Cre	pAdEasy-1
Packaging Cells	293A cells	293A cells
Packaging Strategy	Direct transfection into 293A cells after linearization	Recombination in E. coli after linearization, followed by transfection
Linearization Required for Packaging	No, vector is expressed directly in 293A cells	Yes, linearization after recombination, then transfection into 293A cells
Titer Detection	Plaque assay	Plaque assay
Purification Method	CsCl density gradient centrifugation	CsCl density gradient centrifugation
Advantages	Simple operation, high virus output rate, short cycle	High packaging success rate, low recombination false positives
Disadvantages	Risk of recombination false positives	Complex packaging process, longer cycle

Reference:

Mendonça SA, Lorincz R, Boucher P, et al. Adenoviral vector vaccine platforms in the SARS-CoV-2 pandemic. NPJ Vaccines. 2021;6(1):97.

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