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Flow chart of retrovirus generation and transduction

Retroviral backbone vector features transgenic expression cassette flanked by 5'-LTR and 3'-LTR. Packaged virions can transduce epithelial cells, in this case, membrane GFP expression in MDCK cells.

Custom Retrovirus Production

$1,850.00

Item Cat No.: BCFTRV

Application: Custom Retrovirus Production

Reactivity: Cell culture

BiCell Scientific’s retrovirus production service provides customized retrovirus design and packaging services including retroviral vector cloning, retrovirus generation, packaging and titration, target cell transduction and transgenic expression validation.

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Oncoretroviruses (cancer-causing retroviruses), include human T-lymphotropic virus (HTLV) causing a type of leukemia in humans, and murine leukemia viruses (MLVs) in mice, are powerful tools of gene transfer. The integration of oncoretroviral genome into host cells occurs during cell division, therefore, oncoretrovirus can only transduce dividing cells, but not cells that have entered terminal differentiation, such as neurons.

Oncoretroviruses have gag, pol and env genes, coding for viral proteins in the order: 5´-gag–pol–env-3´. The 5′-end of long terminal repeat (LTR) includes four regions, which are R, U5, PBS (primer binding site), and L; the 3′-end of LTR includes three regions, which are PPT (polypurine tract), U3, and R.

BiCell Scientific’s retrovirus production service provides customized retrovirus design and packaging services including retroviral vector cloning, retrovirus generation, packaging and titration, target cell transduction and transgenic expression validation.

  • Molecular cloning of transgene into retroviral backbone vector
  • Transfection of HEK293T cells with retroviral backbone vector as well as vectors expressing gag–pol and vsv–g
  • Harvesting and purification of retrovirus from cell culture medium
  • Transducing target cells and titrating virus titer with GFP expression
  • Validation of transgene expression in target cells with IF/IHC
  • Microscopic imaging of transgene expression in target cells

Customers will receive the following deliverables at the end of each project:

  • Validated retroviral backbone vector (2nd generation) containing desired transgene
  • Validated pre-packaged retrovirus of 1 ml at titer of 1 x 108 IFU/ml pseudotyped with VSV-G coat protein
  • Validation of target cell transduction at >90% with GFP expression
  • Validation of transgenic expression in target cells with IF/IHC
  • Retroviral vector cloning – 2-3 weeks
  • Retrovirus packaging  – 1-2 weeks
  • Target cell transduction – 1-2 weeks
  • Transgenic expression validation – 2-3 weeks

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"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

School of Medicine | Yale University

"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

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