ALERT

ATTENTION ⚠️

In Observance of U.S. Holiday, Independence Day, Agilent Cross Lab/ iLab Operations Software Support Help Desk will be closed during U.S. hours on Monday July 4th, 2022. We will resume regular U.S. support hours on Tuesday July 5th, 2022. EU and APAC Support will remain open during this time. For urgent matters, please add "Urgent" to the ticket/ email subject or press "1", when prompted, to escalate a call on the iLab Support phone and we will prioritize those requests first.

Virus Vector Core (VVC)

Overview of Services

MISSION

UMB Virus Vector Core (VVC) offers Adeno Associated Virus (AAV) and lentivirus-based gene delivery systems to be used in gene therapy for targeting a gene of interest (GOI) in in-intro and in-vivo studies. The VVC offers services for custom gene cloning and packaging in Adeno Associated Virus or lentivirus systems. The mission of this Core is to provide high-quality viral vectors, reducing the time and cost for the investigators utilizing modern tools and technology. The Core offers free consultancy for gene construct designing strategy, discussing the research project, implementation, and troubleshooting.

SERVICES OFFERED
1. The virus core will generate ready-to-use pre-made viruses (AAVs) that will be available for immediate pick-up.
2. Production of custom vectors – “from scratch” designed according to the needs and specifications of the investigator.
3. A wide selection of serotype and purification grades for in-vivo (live tissue) or in-vitro (primary cell lines).
4. Provision of cloning vectors for insertion of genes of interest prior to vector production.
5. CRISPR-based genome editing, vector cloning, guide RNA designing, and screening in cell lines.
6. The Core will offer consultation service on the design of viral vectors, providing help in choosing virus type and serotype optimal for the cell- and tissue-specific region.

AAV virus

AAV virus is packaged by using triple transfection in HEK393T cells. pHelper plasmid (#112867pAdDeltaF6), red/cap plasmid, and transgene plasmid.

After 72hr of transfection, the virus is purified by either kit or ultracentrifugation method. 

Titer- 10X12 to 10X13 GC/ml, Note- titer may depend on loading size of target gene and serotype.

Quality control

  1. DNA sequencing of target gene
  2. Titer- Titer of the virus tested by qPCR using standard. Titer- 10X12 to 10X13 GC/ml. Note-Titer depends on the 'target gene' Please check with us. 
  3. Purity- AAV samples are routinely inspected by SDS PAGE followed by color staining. This assay confirms to see the AAV capsid protein VP1, VP2, and VP3 molecular weight and relative density.
  4. Endotoxin Free-To avoids the endotoxin-free AAV virus. We used an Endotoxin-free DNA extraction Kit (Qiagen).

Lentivirus

UMB Virus Vector Core generates the lentivirus in 10mm or 150mm plates using a second-generation lentivirus system. We are using standard psPAX2 and envelop protein CMV-SVS-G/ pMD.2G plasmid transfected in the HEK293 cell line. Lentivirus contained clear supernatant or concentrated by PEG is available for the researchers under controlled conditions (in-vitro use).

Ultra-purified lentivirus-Our service provides in-vivo and invitro grade ultra-purified lentivirus. To do this we are using ultracentrifugation with a sucrose cushion.

Lenti Titer- We used Lenti-X™ qRT-PCR Titration Kit (TAKARA)

 

Service                                                                                            Amount/Titer                Internal Price      External Price    Corporate price

Lentivirus

Lentiviral test sample (supernatant for cell culture).                          10ml, 107 – 108 TU/ml           $255.00           $315.00

Lentiviral (supernatant for Cell culture).                                           100ml, 107–108 TU/ml           $820.00           $1015.00

Ultra-purified in-vivo grade concentrated Lentivirus (small scale)       500µl, 108–109 TU/ml            $925.00            $1015.00               $1550

Ultra-purified in-vivo grade concentrated Lentivirus (large scale)        1000µl, 108–109TU/ml          $1215.00          $1415.00               $1950

AAV virus

AAV Customize virus packaging                                                      700-1ml, 1012–1013GC/ml       $1850.00         $2150.00               $2550

AAV stock virus                                                                            100µl,1012–1013 GC/ml            $270.00           $300.00                 $350

 NOTE-

1-All 'internal' and 'external' users please sign in iLab system to pay the bill. 

2-All the above cost subjects are to update every financial year on July 1.

Leadership

Ramesh Chandra Director 410 706 2770 rchandra@som.umaryland.edu

 

Contacts

Name Role Phone Email Location
Ramesh Chandra
Director
 
410 706 2770
 
RChandra@som.umaryland.edu
 
20 Penn Street HSF II, Rm S263 21201