Adenovirus Introduction

Advantages of Recombinant Adenoviral Vectors

1. The genes and structure are researched thoroughly
2. Has the ability to infect most mammalian cell types (both replicative and non-replicative)
3. Accommodates reasonably large transgenes (up to 8 kb)
4. Allows high expression of the recombinant protein
5. May be grown at high titer (more than 10¹¹VP/ml)
6. Is well tolerated, with post-infection viability of the host cells being almost 100%
7. Remains epichromosomal, i.e. does not integrate into the host chromosome so does not inactivate genes or activate oncogenes
8. Can be easily distributed through intravenous injection or take orally
9. Appliable in large scale production and convenient in transportation.

Disadvantages of adenoviral vectors include the following:

(1) expression is transient since the viral DNA does not integrate into the host,

(2) viral proteins are expressed in the adenoviral vector following administration into the host, and

(3) adenoviral vectors are an extremely common human pathogen and in vivo delivery may be hampered by prior host immune response to one type virus.

To overcome the disadvantages mentioned above, Yrbio modified the vector and optimized the production procedure.  Yrbio offer virus production and infection services includes Overexpression service and RNAi service. Yrbio posses sterilized cell culture room (class 2+ biocontainment and above, ISO/DIS 14644-1) which offers Lentiviral handling services, please contact us for details.

1. Recombinant Viral Services for Overexpression 

 Yrbio provide the adenoviral/lentiviral plasmids for the preclinical cell research and animal tests. These products are ready-to-use packaged viruses.

Service Description

1. The recombinant viral services inculdes the plasmid constructing, insert the interest gene and the report genes as needed, and virus preparation.
2. The purified recombinant viruses can reach the titre of·2×10¹⁰- 1×10¹² pfu/ml and 2×10⁸- 2×10⁹ pfu/ml for adenovirus and lentivirus each.
3. Yrbio provides 1-2ml products to the customer.
4. For large scaled production, please contact us for details.

Customer Notices

1. Customer should confirm that the cell lines used can be infected by adenoviruses/lentiviruses. 2. Yrbio does not provide the services to construct tumor gene integrated viruses or other viruses that are harmful to the operators. 3. Customer should provide the details of the target genes (including DNA sequences, NCBI access no. and the MCS,  resistance of the vectors) and the prepared plasmids (>5ug) containing the genes mentioned above. 4. Yrbio will start the services the next day on the arrival of the materials (the delivery time does not include the identificaiton of viruses). 5. Customer is AOR for incorrect material and informations.

Procedure

1. Tool cells and HEK293 cell culture, viral plasmid constructing and preparation;
2. Transfect the tool cells and detect the overexpression of the target genes; 
3. qPCR verification and Western-blot detection (need preliminary tests to optimized the conditions); 
4. Choose the appropriate virus for packaging;
5. Virus verification and  HEK293 cell intermediate culture;
6. Virus amplification and purification,
7. titre assay, qPCR test and Western-blot detection;
8. Data processing the turn over the reports.

Service Commitment

• 1.  Yrbio provide 20μg viral plasmid for overexpression and 500μl corresponding glycerin bacteria; 
• 2.  Yrbio provide 2*10⁸TU/ml Lentiviruses or 1*10¹¹TU/ml adenovirus; 
• 3.  Transfer the reports in detail, include the procedure, instruments, protocols, raw data and images.

Delivery Time

Lot No.	Contents	Delivery Time (day)	Price (￥)
	Antibody Preliminary Test (optional)	10	1000
	qPCR Preliminary Test (optional)	15	1000
	Plasmid Construction	see also the Plasmid Construction Services
	Plasmid Preparation and Verification (optional for sequencing)	5 (without sequencing delays)	400（100μg）
	HEK293 preparation and  culture	5	2000
	Viral Packaging	15	4000
	Viral Verification	15	1500
	Viral Aplification	20-30	6000
	qPCR titre assay +WB expression detection	20	1500 (qPCR) 1500/membrane
	Data processing	2	Free

2. Recombinant Viral Vectors for RNAi

Yrbio provide the adenoviral/lentiviral plasmids for the preclinical cell research and animal tests. These products are ready-to-use packaged viruses.·

Service Description

1. The recombinant viral services inculdes the plasmid constructing, insert the shRNA fragments and the report genes as needed, and virus preparation.
2. Constructing the controllable expression plasmids (optional).
3. The purified recombinant viruses can reach the titre of·2×10¹⁰- 1×10¹² pfu/ml and 2×10⁸- 2×10⁹ pfu/ml for adenovirus and lentivirus each.
4. Yrbio provides 1-2ml products to the customer.
5. For large scaled production, please contact us for details.

Customer Notices

1. Customer should provide the details of the target genes (including the DNA sequence, NCBI access no. and expression level in the target cells)
2. Yrbio will following the sequence to design 3-4 RNAi target sequences and than construct the plasmid and verified (sequencing) followed by virus packaging as described above
3. If the customer provide the shRNA or RNAi target sequences, the detailed informations of the shRNA should be companioned. 
4. Yrbio will start the services on the next day on the arrival of the materials (the delivery time does not include the identificaiton of viruses).
5. Customer is AOR for incorrect material and informations.

Procedure

1. Tool cells and·HEK293 cell culture, viral plasmid contructing and preparation;
2. Transfect the tool cells and detect the overexpression of the target genes;·
3. qPCR verification and Western-blot·detection (need preliminary tests to optimized the conditions);·
4. Choose the appropriate virus for packaging;
5. Virus verification and ·HEK293 cell intermediate culture;
6. Virus amplification and purification;·
7. titre assay, qPCR test and Western-blot·detection;
8. Data processing the turn over the·reports.

Service Commitment 

• 1. ·Yrbio provide 20μg viral plasmid for overexpression and 500μl corresponding glycerin bacteria;·
• 2. ·Yrbio provide 2*10⁸TU/ml Lentiviruses or 1*10¹¹TU/ml adenovirus;·
• 3. ·Transfer the reports in detail, include the procedure, instruments, protocols, raw data and images.

Delivery Time

Lot No.	Contents	Delivery Time (day)	Price (￥)
	Antibody Preliminary Test (optional)	10	1000
	qPCR Preliminary Test (optional)	15	1000
	Plasmid Construction	see also the Plasmid Construction Services
	Plasmid Preparation and Verification (optional for sequencing)·	5 (without sequencing delays)	400（100μg）
	HEK293 preparation and ·culture	5	2000
	Viral Packaging	15	4000
	Viral Verification	15	1500
	Viral Aplification	20-30	6000
	qPCR titre assay +WB expression detection	20	1500 (qPCR) 1500/membrane
	Data processing	2	Free