AAV (Adenoassociated Virus) Packaging Services

Adeno-associated Virus

Adeno-associated viruses (AAVs) belong to the Parvoviridae family, comprising a group of small-sized non-enveloped viruses with an icosahedral capsid carrying a single-stranded DNA genome of approximately 4.7 kilobases (kb). AAV is one of the most popular gene delivery vehicles due to its multiple, wide-ranging tropism profiles, low immunogenicity, long-term stable gene expression, and strong diffusion capabilities.

At OBiO, we can supply you a holistic vector design, gene synthesis, plasmid and AAV production services. From idea to AAV, we can help you to delivery gene to any kind of cell you want to target, both in vivo and in vitro.

* We offer a variety of packaging systems for AAV vectors, in addition to the traditional Triple Plasmid System, we can also provide the dual plasmid system and sf9-baculovirus for Production of AAV, and we offer large scale AAV preps are produced in cell factory or shake flask.

*Available purification method: ultracentrifugation, chromatography

Types of AAV we offered

*Single-stranded AAV(ssAAV) and self-complementary AAV(scAAV).

*100+ different serotypes available：AAV1-9, AAV-PHP.eB, AAV-PHP.S, AAV2-retro, AAV-ie, AAV-LungM3, AAV-LungX, AAV-GC01, AAV-GC02, AAV-GC03, AAV-GC13, AAV-DJ, AAV2.7m8, AAV6.2ff, AAV-PAN, AAVRec2, AAVrh10, custom AAV serotype

Quality, scale and turnaround time of AAV

OBiO provides flexible scaling options. Our high titer AAV packaging services guarantee titers ≥1E+13vg/mL. For advanced translational research, our NHP grade AAV production ensures an empty capsid rate of <10% and strict purity control.

	Research Grade	NHP Grade
Suitable for	Rodents, other small mammals	Dog, Pig, Sheep, NHP, other larger animals
Quality standard	Titer ≥ 1E+13vg/mL （with ddPCR/qPCR） AAV Capsid determination：Match capsid protein size Sterility test：Negative Endotoxin Assay（optional）：＜10EU/mL Mycoplasma Detection（optional）：Negative	Titer ≥1E+13vg/mL（with ddPCR） AAV Capsid determination：Match capsid protein size Residual Nuclease：＜0.1ng/ml HCP：＜20ng/ml Purity Assay：＞95% Endotoxin Assay：＜10EU/mL Empty Capsid Rate：＜10% Sterility test：Negative Mycoplasma Detection：Negative
Scale	multiple scales from1E+12vg to 5E+13vg	multiple scales from1E+13vg to 1E+15vg

* Guaranteed titers are suit for regular packaging, in which, (1) the payload gene size cannot exceed the AAV packaging capacity (4.7 kb from ITR to ITR), (2) you can choose AAV1, AAV5, AAV8, AAV9, AAV-retro, AAV-PHP.eB, AAV-PHP.S, AAV-ie, AAV-LungM3, AAV-LungX, AAV-DJ, AAV2.7m8, AAV-PAN, AAVRec2 or AAVrh10 to package your GOI.

* For big size gene packaging or low-yield AAV serotypes, or novel AAV, we can supply you the virus with typical titer > 5E+12 vg/mL, any needs could be inquired.

* For NHP grade AAV producing, we can supply you AAV manufacturing with multi-serotypes, any needs could be inquired.

* Turnaround time depends on the complexity of gene synthesis, vector construction and kind of AAV serotype. Listed time is an estimated time. We will report you a confirmed time based on the technical proposal.

Tissue-tropism of adeno-associated (AAV) viral vectors

Choosing the right serotype is critical for AAV gene delivery. Below is our AAV tissue tropism chart.

Serotype	Tissue tropism
AAV1	CNS, skeletal muscle, smooth muscle, retina, Endothelial Cell
AAV2	CNS, retina, liver, inner ear, kidney, smooth muscle
AAV3	muscle, liver, lung
AAV4	CNS, retina, muscle
AAV5	CNS, smooth muscle, retina, lung
AAV6	lung, CNS, smooth muscle, heart
AAV7	muscle, liver
AAV8	CNS, liver, kidney, adipose, retina, muscle, pancreas
AAV9	CNS, heart, lung, retina, skin
AAVDJ	retina, liver, kidney, in-vitro
AAV2-Retro	CNS：retrograde tracers
AAV9-PHP.B	CNS：crossed the blood-brain barrier
AAV9-PHP.eB	CNS：crossed the blood-brain barrier
AAV9-PHP.S	PNS：intravenous administration
AAV2-7m8	retina
AAV-LungX	pulmonary vascular endothelial cells
AAV-LungM3	lung
AAV-PAN	pancreas（intraperitoneal injection）
Anc80L65	inner ear, retina, skeletal muscle,liver
AAV6-ShH10Y	muller cells
AAV2-BR1	brain microvasculature endothelial cell
AAVRec2	adipose
AAV6-TM6	microglia
AAV6.2FF	lung, muscle
AAV2-rh10	CNS, liver, heart, lung, in-vitro
AAV-DSS	bone
AAV9-MyoAAV 1A	skeletal muscle, Cardiac muscle
AAV-Vec	vascular endothelial cells
AAV-ie	inner ear
AAV-ie-K558R	inner ear
AAV-DS1	inner ear
AAV-BI30	CNS: Endothelial Cells
AAV-MaCPNS1	PNS in rodent, PNS and CNS in macaque and marmoset (systemic administration)
AAV-MaCPNS2
AAV-MG1.2	microglia with high efficiency
AAV-GC01	retina：the retinal pigment epithelium (intravitreal injection)
AAV-GC02	retina：the retinal pigment epithelium (intravitreal injection)
AAV-GC03	retina (intravitreal injection)
AAV-GC13	retina: Cone and Rod (intravitreal injection)

* Please note, virus genome backbone is from AAV2, i.e. we use ITR2 for packaging.

Different serotypes are distinguished by the different capsid proteins.

Promoter choices

For cell-specific gene expression, listed promoters can be designed into your vector.

Organ	Name	Cell Type Specificity
Brain/CNS	SYN1	Mature neurons
	CaMKIIa	Pyramidal neurons
	TH	Dopaminergic neurons
	GFAP/GfaABC1D	Astrocytes
	CX3CR1	Microglia
	Iba1	Microglia
	F4/80	Microglia
	GAD67	GABAergic neurons
	VGAT1	GABAergic neurons
	mDLX	GABAergic neurons
	fPV	Inhibitory neuron subtype (PV)
	fSST	Inhibitory neuron subtype (SST)
	fNPY	Inhibitory neuron subtype (NPY)
	DAT	Dopaminergic neurons
	L7pcp2	Purkinje cells
	MBP	oligodendrocytes
	NG2/NGL2	NG2-glia/oligodendrocyte precursor cells
	PRSx8	adrenergic neurons/NE neurons
	ChAT	cholinergic neurons
	HCRT	Hcrt neurons
	FEV	serotonergic neurons
	Nestin	Neural stem cells (NSCs)
Retina	ProA1	Cone photoreceptor
	hRHO	Rod photoreceptor
	BEST1（VMD2）	RPE
Inner cells	IHC	Inner hair cells
	OHC	Outer hair cells
	HC	Hair cells
	OPC	Outer pillar cells
	IBC	Inner border cells
	SC1	Supporting cells
	SGN I	Type I neuron
	SGN II	Type II neuron
Mammalian Inducible Promoters	TRE	tetracycline-inducible promoters
Mammalian Inducible Promoters	c-FOS	c-fos+ neurons/engram cells
Heart	cTnTo/cTnT	Cardiomyocytes
	TCF21	Cardiac fibroblasts
	Postn	Cardiac myofibroblast
Liver	Alb	Mature hepatocytes
	TBG/Apoe	Hepatocytes
	GFAP/α-SMA	Hepatic stellate cells
	F4/80	Kupffer cells
Lung	SP-C	AT II cells (alveolar type II epithelial cells)
	SP-B	AT II cells and Clara cells (bronchiolar epithelial cells)
	CC10	Bronchial epithelial cells
Adipose	Adipoq	Adipocytes
Muscle	MHCK7	Differentiated postmitotic striated muscle cells
	SM22a	Vascular smooth muscle cells
	dMCK/tMCK	skeletal muscles
	Myog	Myoblasts
Intestine	Villin	Intestinal epithelial cells
Intestine	MUC2	Intestinal goblet cells
Pancreas	HIP/Mlp2/Ins2/Pdx1	Islet β-cells
Pancreas	GCG	Islet α-cells
ECs	hFLT1	Vascular endothelial cells
ECs	Tie1	Endothelial cells
Mammalian Ubiquitous Promoters	CMV	Strong promoter; may have variable strength in some cell types.
	EF1A	Strong promoter.
	EF1	Medium-strength promoter
	EFS	Medium-strength promoter
	CAG	Strong promoter
	CBh	Strong promoter
	SFFV	Strong promoter; drives high levels of gene expression, particularly in cell types of the myeloid lineage.
	mPGK	Medium-strength promoter
	hPGK	Medium-strength promoter
	UBC	Weak promoter

FAQs of Adeno-associated Virus

What is the packaging capacity of your custom AAV vectors?

A: The standard AAV packaging capacity is approximately 4.7 kb (from ITR to ITR). For larger gene payloads, please contact our technical team for custom vector design solutions.

How do you determine the empty capsid rate for NHP-grade AAV?

A: For our NHP-grade AAV packaging services, we perform stringent quality control including Cryo-EM or AUC assays to guarantee an empty capsid rate of less than 10%.

What is the notation form of AAV serotypes?

The commonly used rAAV in research is a recombinant viral vector produced by combining the AAV2 genome with different AAV capsids, denoted as rAAV2/N (where N represents different serotypes, such as the common rAAV2/8, rAAV2/9, etc.).

What's the cloning capacity for rAAVs?

The payload capacity of rAAV (the size of the fragment between the two ITRs) is 4.7kb.

AAV has a packaging capacity of ~4.7Kb (from ITR to ITR). Since the two ITRs of AAV are about ~0.2Kb total, the foreign DNA (promoter + GOI + polyA + WPRE, etc.) that can be introduced between these two ITRs should be smaller than 4.5Kb. When the length of inserted DNA between the two ITRs is close to the maximum allowed, the packaging efficiency decreases significantly. OBiO TECH has optimized and shortened sequences in the promoter/polyA/WPRE, which increases the AAV payload capacity.

For self-complementary AAV (scAAV), the capacity is half that of the single-stranded AAV (ssAAV).

What is the difference of AAV titer unit between GC/mL and vg/mL?

In the context of AAV titer, "GC/mL" (genome copies per mL) and "vg/mL" (vector genomes per mL) are essentially interchangeable terms, both units represent the number of viral genomes present in a given volume of solution. AAV titer measuring method is using qPCR to detect the exogenous DNA copies in virus genome.

What is the optimal time for AAV expression?

The optimal time for AAV expression typically occurs several days to a few weeks post-injection. For in vivo studies, single-stranded AAV is recommended to be detected 3 weeks post injection, and scAAV is detected one week post injection. For in vitro, it is suggested to detect 2-3 days post infection.

Can AAV be used to infect cells in vitro?

Yes. AAV can be used in vitro. Due to the different tropism of AAV serotypes for various cells, a suitable MOI can be obtained through pre-infection experiments before use.