Viral Vector Manufacturing

VectorBuilder has extensive expertise in producing various types of viral vectors for cell and gene therapy development. We offer these at various scales and quality attributes to meet the full range of demands along the gene therapy drug development pipeline. Our team has established and validated platform technologies for large-scale GMP manufacturing of adeno-associated virus (AAV) and lentivirus, and we also have the expertise and capacity for manufacturing other types of viral vectors such as adenovirus, MMLV, herpes simplex virus (HSV), and vesicular stomatitis virus (VSV).

Highlights

Experts in Virus Packaging Proprietary technologies optimized to achieve the highest titer and potency at any scale.

Scalable Solutions Versatile platform to support your production needs, from research to commercial grades.

Highest Quality and Consistency State-of-the-art GMP facilities and equipment providing the highest purity and consistency.

Grades of Viral Vectors Offered

We produce viruses in several grades to cover a full range of downstream applications including drug discovery research, preclinical studies, clinical trials, and commercialization.

R&D

Preclinical

Clinical

Commercial

Research-Grade

GMP-Like

GMP-Grade

Suitable for basic research, such as studies in gene function and regulation, and drug discovery studies
Produced in conventional cell culture facilities with stringent QC

Suitable for preclinical studies (e.g. animal testing of drug safety and metabolism)
Produced in PAS or GMP facilities in a manner that adopts key features of GMP guidelines
Can be produced under antibiotic-free and animal component-free conditions

Suitable for preclinical studies, clinical studies, and commercialization
Produced in our certified GMP suite with strict adherence to GMP guidelines
Includes comprehensive and customizable QC assays and batch release report

Comparison of different grades of virus

AAV

Lentivirus

	Research-Grade	GMP-Like	GMP-Grade
Applications	Basic research, drug discovery, and preclinical studies	Preclinical studies such as animal testing of drug safety and metabolism	Preclinical studies, clinical studies, and commercialization
Production scales	5x10¹⁰to 10¹⁴ GC per batch	5x10¹³to 10¹⁷ GC per batch	10¹⁴to 10¹⁷ GC per batch
Turnaround time	10-50 days	4-5 months	6-12 months
Production site	In parallel production in standard BSL-2 laboratory	Produced in segregated BSL-2 production suites	Produced in certified GMP suites (BSL-2)
Quality system	ISO9001	ISO9001 while adopting key features of GMP manufacturing	ICH quality guidelines for GMP manufacturing
Document control and traceability	No	Yes	Full traceability
Vector characterization	RE digestion, insert sequencing	RE digestion, full-length sequencing	RE digestion, full-length sequencing
Process development	No	Performed on a case-by-case basis depending on individual project needs	Yes
Cell banking	No	Available upon request	Yes
Antibiotic-free	No	Available upon request	Yes
Animal component-free	No	Available upon request for suspension culture	For suspension culture
Purification	Ultracentrifugation	Ultracentrifugation, affinity purification, IEX purification, mixed-mode chromatography	Ultracentrifugation, affinity purification, IEX purification, mixed-mode chromatography
QC and release tests	Titer measurement, SDS-PAGE, endotoxin detection, sterility testing, mycoplasma detection, etc.	Performed on a case-by-case basis depending on individual project needs	Full panel QC assays, analytical development upon individual project needs
Aseptic fill/finish	No	Available upon request	Yes
Storage of retain sample	Available upon request	Available upon request	Yes
Document deliverable	COA	1. COA 2. Manufacturing summary 3. TSE/BSE statement upon request	1. COA 2. TSE/BSE statement 3. CTD documents 4. Others (BMR etc.) upon request

	Research-Grade	GMP-Like	GMP-Grade
Applications	Basic research, drug discovery, preclinical studies	Preclinical studies such as animal testing of drug safety and metabolism	Preclinical studies, clinical studies, and commercialization
Production scales	>2.5x10⁷ TU	10⁹ to 10¹² TU per batch	5x10⁹ to 10¹² TU per batch
Turnaround time	8-16 days	4-5 months	6-12 months
Production site	In parallel production in standard BSL-2 laboratory	Produced in segregated BSL-2 production suites	Produced in certified GMP suites (BSL-2)
Quality system	ISO9001	ISO9001 while adopting key features of GMP manufacturing	ICH quality guidelines for GMP manufacturing
Document control and traceability	No	Yes	Full traceability
Vector characterization	RE digestion, insert sequencing	RE digestion, full-length sequencing	RE digestion, full-length sequencing
Process development	No	Performed on a case-by-case basis depending on individual project needs	Yes
Cell banking	No	Available upon request	Yes
Antibiotic-free	No	Available upon request	Yes
Animal component-free	No	Available upon request for suspension culture	For suspension culture
Purification	Ultracentrifugation	Ultracentrifugation, affinity purification, IEX purification, mixed-mode chromatography	Ultracentrifugation, affinity purification, IEX purification, mixed-mode chromatography
QC and release tests	Titer measurement, sterility testing, mycoplasma detection, etc.	Performed on a case-by-case basis depending on individual project needs	Full panel QC assays, analytical development upon individual project needs
Aseptic fill/finish	No	Available upon request	Yes
Storage of retain sample	Available upon request	Available upon request	Yes
Document deliverable	COA	1. COA 2. Manufacturing summary 3. TSE/BSE statement upon request	1. COA 2. TSE/BSE statement 3. CTD documents 4. Others (BMR etc.) upon request

Workflow for GMP Production of Viral Vectors

VectorBuilder strives to deliver products of the highest quality on time. Here is a typical GMP production workflow of viral vectors:

Vector
Characterization

Process
Development

Cell
Banking

Plasmid
Production

Engineering
Run

GMP Run

QC & Release

1 week

2.5 months

1 month

RE digestion
Sequencing

Cell line
development
USP/DSP
Analytical
development

Segregated
suite
(GMP-like)
GMP suite
(GMP-grade)

Closed system
Fill/finish

General/physical
properties
Identity
Purity
Safety

Platform Technologies

AAV Platform

Lentivirus Platform

AAVs are packaged in HEK293 cells under either adherent conditions (multi-layer flasks or fixed-bed bioreactors) or serum-free suspension conditions (up to 200 L single-use bioreactors). Upon request, we can also package AAV in suspension Sf9 insect cells. We are capable of production scales reaching up to 10¹⁷ GC AAV per batch.

Cell Bank recovery and culture > Fermentation > Harvest and lysis > Centrifugation > Chromatography > TFF > Sterile filtration > QC

Adherent
(Multi-layer flasks,
fixed-bed bioreactors)

Suspension
(Single-use bioreactors)

Transfection
(PEI, calcium
phosphate

Harvest and
lysis

Clarification and
concentration
(TFF)

Purification
(Chromatography)

Formulation
(TFF, sterile
filtration, fill/finish)

QC and release

Lentivirus packaging (2nd or 3rd generation, pseudotyped with VSV-G or other viral surface proteins) is performed in HEK293 cells under either adherent conditions (multi-layer flasks or fixed-bed bioreactors) or serum-free suspension conditions (up to 200 L single-use bioreactors). We are capable of production scales of up to 10¹² TU per batch.

Cell Bank recovery and culture > Fermentation > Harvest and alkaline lysis > Centrifugation > Chromatography > TFF > Sterile filtration > QC

Adherent
(Multi-layer flasks,
fixed-bed bioreactors)

Suspension
(Single-use bioreactors)

Transfection
(PEI, calcium
phosphate

Clarification and
concentration
(TFF)

Purification
(Chromatography)

Formulation
(TFF, sterile
filtration, fill/finish)

QC and release

For more information on our manufacturing platform and/or GMP capabilities, contact the VectorBuilder service team.

Talk to Our Experts

Analytical Methods

Quality control assays

The table below summarizes the default QC assays (marked with ✓) for the release of GMP-like and GMP-grade viral vectors. Optional QC assays are performed depending on individual project needs.

AAV Platform

Lentivirus Platform

Attribute		QC Assay	GMP-Like	GMP-Grade
General/Physical Properties	Osmolality	Freezing point depression	Optional	✓
	pH	pH meter	✓	✓
	Particulate matter	Light obscuration particle count test	Optional	✓
	Appearance	Visual inspection	Optional	✓
	Extractable volume	Direct measurement	Optional	✓
	Physical titer	qPCR	✓	✓
		ddPCR	Optional	✓
		ELISA	✓	✓
	Infectious titer	TCID50	Optional	Optional
Potency	mRNA transcription	RT-qPCR	Optional	✓
	mRNA transcription	RT-ddPCR	Optional	Optional
	Protein expression	Flow cytometry (FCM)	Optional	Optional
Identity	Full-length plasmid sequence	Sanger sequencing	✓	✓
		NGS	Optional	Optional
		Third-generation sequencing	Optional	Optional
Purity	Capsid protein purity	SDS-PAGE with silver staining	✓	✓
	Empty capsid ratio	Transmission electron microscopy (TEM)	Optional	Optional
		Mass photometry (MP)	✓	✓
		Analytical ultracentrifugation (AUC)	Optional	✓
		Charge detection mass spectrometry (CDMS)	Optional	Optional
	Aggregation	Dynamic light scattering (DLS)	Optional	Optional
	Aggregation	Size exclusion chromatography (SEC)	Optional	✓
	Genome integrity	Alkaline gel electrophoresis	✓	✓
Process-Related Impurity	Residual plasmid DNA	qPCR	✓	✓
	Residual plasmid DNA	ddPCR	Optional	Optional
	Residual host cell DNA	qPCR	✓	✓
	Residual E1A sequence	qPCR	✓	✓
	Residual host cell DNA fragment size	Capillary electrophoresis (CE)	Optional	✓
	Residual host cell proteins	ELISA	✓	✓
	Residual Benzonase	ELISA	✓	✓
	Residual affinity ligand	HPLC	Optional	Optional
	Residual transfection reagent	HPLC	Optional	Optional
	Residual ultracentrifugation agent	HPLC	Optional	✓
	Residual detergent	HPLC	Optional	Optional
Product-Related Impurity	Replication competent AAV	qPCR	Optional	✓
Safety	Sterility	Direct inoculation	Optional	Optional
	Sterility	Membrane filtration sterility test	✓	✓
	Endotoxin	Kinetic chromogenic assay (KCA)	✓	✓
	Mycoplasma	DNA staining	✓	Optional
		qPCR	✓	Optional
		Direct culture and indicator cell culture	Optional	✓
	Adventitious virus testing	Cell culture (using MRC-5, VERO, and HEK293 cells)	Optional	✓
Documentation	-	-	1. COA 2. Manufacturing summary 3. TSE/BSE certificate upon request	1. COA 2. TSE/BSE certificate 3. CTD documents 4. Others (BMR, etc.) upon request

Attribute		QC Assay	GMP-Like	GMP-Grade
General/Physical Properties	Osmolality	Freezing point depression	Optional	✓
	pH	pH meter	✓	✓
	Particulate matter	Light obscuration particle count test	Optional	✓
	Appearance	Visual inspection	Optional	✓
	Extractable volume	Direct measurement	Optional	✓
	Physical titer	RT-qPCR	Optional	✓
	Physical titer	ELISA	✓	✓
	Functional titer	qPCR	✓	✓
	Functional titer	Flow cytometry (FCM)	Optional	✓
	Infectious titer	TCID50	Optional	Optional
Potency	mRNA transcription	RT-ddPCR	Optional	Optional
Potency	Protein expression	Flow cytometry (FCM)	Optional	Optional
Identity	Full-length plasmid sequence	Sanger sequencing	✓	✓
		NGS	Optional	Optional
		Third-generation sequencing	Optional	Optional
Purity	Aggregation	Dynamic light scattering (DLS)	Optional	Optional
Process-Related Impurity	Residual plasmid DNA	qPCR	✓	✓
	Residual plasmid DNA	ddPCR	Optional	Optional
	Residual host cell DNA	qPCR	✓	✓
	Residual E1A sequence	qPCR	✓	✓
	Residual SV40 LTA sequence	qPCR	✓	✓
	Residual host cell DNA fragment size	Capillary electrophoresis (CE)	Optional	✓
	Residual host cell proteins	ELISA	✓	✓
	Residual Benzonase	ELISA	✓	✓
	Residual affinity ligand	HPLC	Optional	Optional
	Residual transfection reagent	HPLC	Optional	Optional
	Residual detergent	HPLC	Optional	Optional
Product-Related Impurity	Replication competent lentivirus	Cell culture	Optional	✓
Safety	Sterility	Direct inoculation	Optional	Optional
	Sterility	Membrane filtration sterility test	✓	✓
	Endotoxin	Kinetic chromogenic assay (KCA)	✓	✓
	Mycoplasma	DNA staining	✓	Optional
		qPCR	✓	Optional
		Direct culture and indicator cell culture	Optional	✓
	Adventitious virus testing	Cell culture (using MRC-5, VERO, and HEK293 cells)	Optional	✓
Documentation	-	-	1. COA 2.Manufacturing summary 3.TSE/BSE certificate upon request	1. COA 2. TSE/BSE certificate 3. CTD documents 4. Others (BMR, etc.) upon request

Additional QC services not listed above may be provided upon request. Please contact our service team for more details.

Talk to Our Experts

Stability studies

Drug stability studies are performed following ICH guidelines. Forced degradation studies are performed to evaluate the influence of light, temperature, pH, etc., on the quality attributes of the viral vector DNA.

Our Facilities

VectorBuilder currently has about 100,000 ft² of modern GMP-compliant facilities with advanced designs and state-of-the-art equipment meeting GMP regulations of US, EU, Japan, China, and PIC/S. Our facilities comply with ICH guidelines and are suitable for GMP production for Phase I/II/III clinical trials.

Process & Analytical
Development Suites
GMP Manufacturing
Suites
Fill/finish Suites
QC Laboratories
GMP Warehouse

We have multiple GMP suites totaling 8,400 ft² for PD/pilot runs. All BSL-2 certified areas are set up with Grade A BSC in Grade C environment.

Our suites are designed for manufacturing plasmid, viral vector, and cell line production at various scales. Each suite is equipped with independent airflow. Our suites contain Grade A BSC in Grade B/C environment and are BSL-2 certified.