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FreeStyle 293-F



Homo sapiens


Transformant: NCBI_TaxID; 28285; Adenovirus 5. Discontinued: ATCC; PTA-5080. DT Created: 22-10-12; Last updated: 05-07-19; Version: 13

Proper Citation

ATCC Cat# PTA-5080, RRID:CVCL_D603


Patent:US7217566; DT Created: 22-10-12; Last updated: 05-07-19; Version: 13


Transformed cell line DT Created: 22-10-12; Last updated: 05-07-19; Version: 13


DT Created: 22-10-12; Last updated: 05-07-19; Version: 13


Freestyle 293, Freestyle 93F, Freestyle HEK293F, 293-F FreeStyle DT Created: 22-10-12, Last updated: 05-07-19, Version: 13



Cat Num


Cross References

BTO; BTO:0005267 ATCC; PTA-5080 FCS-free; 38-2-44-1-8-3 Lonza; 885 Wikidata; Q54835158 DT Created: 22-10-12; Last updated: 05-07-19; Version: 13


DT Created: 22-10-12; Last updated: 05-07-19; Version: 13

Originate from Same Individual

DT Created: 22-10-12; Last updated: 05-07-19; Version: 13

Engineered Dengue Virus Domain III Proteins Elicit Cross-neutralizing Antibody Responses in Mice.

  • Frei JC
  • J. Virol.
  • 2018 Jul 5

Literature context:


Dengue virus is the most globally prevalent mosquito-transmitted virus. Primary infection with one of four co-circulating serotypes (DENV1-4) causes a febrile illness but secondary infection with a heterologous serotype can result in severe disease due in part to antibody dependent enhancement of infection (ADE). In ADE, cross-reactive but non-neutralizing antibodies, or sub-protective levels of neutralizing antibodies, promote uptake of antibody-opsonized virus in Fc-γ receptor-positive cells. Thus, elicitation of broadly neutralizing antibodies (bNAbs), but not non-neutralizing antibodies, is desirable for Dengue vaccine development. Domain III of the envelope glycoprotein (EDIII) is targeted by bNAbs and thus an attractive immunogen. However, immunization with EDIII results in sera with limited neutralization breadth. We developed "resurfaced" EDIII immunogens ("rsDIIIs") whereby the A/G-strand epitope that is targeted by bNAb 4E11 is maintained, but less desirable epitopes are masked. RsDIIIs bound 4E11, but not serotype-specific or non-neutralizing antibodies. One rsDIII and, unexpectedly, WT DENV-2 EDIII elicited cross-neutralizing antibody responses against DENV1-3 in mice. While these sera were cross-neutralizing, they were not sufficiently potent to protect AG129 immunocompromised mice at a dose of 200 μL (FRNT50 at titer of ∼1:60-1:130) against mouse adapted DENV-2. Our results provide insight into immunogen design strategies based on EDIII.IMPORTANCE Dengue virus causes approximately 390 million infections per year. Primary infection by one serotype causes a self-limiting febrile illness, but secondary infection by a heterologous serotype can result in Severe Dengue, which is characterized by hemorrhagic fever and shock syndrome. This severe disease is thought to arise because of cross-reactive, non- or poorly-neutralizing antibodies from the primary infection, that are present in serum at the time of secondary infection. These cross-reactive antibodies enhance the infection rather than controlling it. Therefore, induction of a broadly and potently neutralizing antibody response is desirable for Dengue vaccine development. Here we explore a novel strategy developing immunogens based on domain III of the E glycoprotein, where undesirable epitopes (non-neutralizing, or non-conserved) are masked by mutation. This work provides fundamental insight into the immune response to domain III that can be leveraged for future immunogen design.

Funding information:
  • Biotechnology and Biological Sciences Research Council - (United Kingdom)
  • NIAID NIH HHS - P01 AI106695(United States)
  • NIAID NIH HHS - R01 AI075647(United States)
  • NIAID NIH HHS - R21 AI128090(United States)
  • NIGMS NIH HHS - T32 GM007491(United States)

Visualization of ligand-induced transmembrane signaling in the full-length human insulin receptor.

  • Gutmann T
  • J. Cell Biol.
  • 2018 May 7

Literature context:


Insulin receptor (IR) signaling plays a critical role in the regulation of metabolism and growth in multicellular organisms. IRs are unique among receptor tyrosine kinases in that they exist exclusively as covalent (αβ)2 homodimers at the cell surface. Transmembrane signaling by the IR can therefore not be based on ligand-induced dimerization as such but must involve structural changes within the existing receptor dimer. In this study, using glycosylated full-length human IR reconstituted into lipid nanodiscs, we show by single-particle electron microscopy that insulin binding to the dimeric receptor converts its ectodomain from an inverted U-shaped conformation to a T-shaped conformation. This structural rearrangement of the ectodomain propagates to the transmembrane domains, which are well separated in the inactive conformation but come close together upon insulin binding, facilitating autophosphorylation of the cytoplasmic kinase domains.

Funding information:
  • NCI NIH HHS - R01CA155413(United States)

The Marburgvirus-Neutralizing Human Monoclonal Antibody MR191 Targets a Conserved Site to Block Virus Receptor Binding.

  • King LB
  • Cell Host Microbe
  • 2018 Jan 10

Literature context:


Since their first identification 50 years ago, marburgviruses have emerged several times, with 83%-90% lethality in the largest outbreaks. Although no vaccines or therapeutics are available for human use, the human antibody MR191 provides complete protection in non-human primates when delivered several days after inoculation of a lethal marburgvirus dose. The detailed neutralization mechanism of MR191 remains outstanding. Here we present a 3.2 Å crystal structure of MR191 complexed with a trimeric marburgvirus surface glycoprotein (GP). MR191 neutralizes by occupying the conserved receptor-binding site and competing with the host receptor Niemann-Pick C1. The structure illuminates previously disordered regions of GP including the stalk, fusion loop, CX6CC switch, and an N-terminal region of GP2 that wraps about the outside of GP1 to anchor a marburgvirus-specific "wing" antibody epitope. Virus escape mutations mapped far outside the MR191 receptor-binding site footprint suggest a role for these other regions in the GP quaternary structure.

Funding information:
  • Canadian Institutes of Health Research - (Canada)
  • NIAID NIH HHS - R01 AI089498()
  • NIAID NIH HHS - R21 AI121799()
  • NIAID NIH HHS - U19 AI109711()
  • NIAID NIH HHS - U19 AI109762()
  • NIGMS NIH HHS - P41 GM103393()

HIV Envelope Glycoform Heterogeneity and Localized Diversity Govern the Initiation and Maturation of a V2 Apex Broadly Neutralizing Antibody Lineage.

  • Landais E
  • Immunity
  • 2017 Nov 21

Literature context:


Understanding how broadly neutralizing antibodies (bnAbs) to HIV envelope (Env) develop during natural infection can help guide the rational design of an HIV vaccine. Here, we described a bnAb lineage targeting the Env V2 apex and the Ab-Env co-evolution that led to development of neutralization breadth. The lineage Abs bore an anionic heavy chain complementarity-determining region 3 (CDRH3) of 25 amino acids, among the shortest known for this class of Abs, and achieved breadth with only 10% nucleotide somatic hypermutation and no insertions or deletions. The data suggested a role for Env glycoform heterogeneity in the activation of the lineage germline B cell. Finally, we showed that localized diversity at key V2 epitope residues drove bnAb maturation toward breadth, mirroring the Env evolution pattern described for another donor who developed V2-apex targeting bnAbs. Overall, these findings suggest potential strategies for vaccine approaches based on germline-targeting and serial immunogen design.

Funding information:
  • NHLBI NIH HHS - R21-HL122443(United States)

Identification of GPC2 as an Oncoprotein and Candidate Immunotherapeutic Target in High-Risk Neuroblastoma.

  • Bosse KR
  • Cancer Cell
  • 2017 Sep 11

Literature context:


We developed an RNA-sequencing-based pipeline to discover differentially expressed cell-surface molecules in neuroblastoma that meet criteria for optimal immunotherapeutic target safety and efficacy. Here, we show that GPC2 is a strong candidate immunotherapeutic target in this childhood cancer. We demonstrate high GPC2 expression in neuroblastoma due to MYCN transcriptional activation and/or somatic gain of the GPC2 locus. We confirm GPC2 to be highly expressed on most neuroblastomas, but not detectable at appreciable levels in normal childhood tissues. In addition, we demonstrate that GPC2 is required for neuroblastoma proliferation. Finally, we develop a GPC2-directed antibody-drug conjugate that is potently cytotoxic to GPC2-expressing neuroblastoma cells. Collectively, these findings validate GPC2 as a non-mutated neuroblastoma oncoprotein and candidate immunotherapeutic target.

Funding information:
  • NCI NIH HHS - T32 CA009615()
  • NIGMS NIH HHS - T32 GM008638()

A potent human neutralizing antibody Fc-dependently reduces established HBV infections.

  • Li D
  • Elife
  • 2017 Sep 26

Literature context:


Hepatitis B virus (HBV) infection is a major global health problem. Currently-available therapies are ineffective in curing chronic HBV infection. HBV and its satellite hepatitis D virus (HDV) infect hepatocytes via binding of the preS1 domain of its large envelope protein to sodium taurocholate cotransporting polypeptide (NTCP). Here, we developed novel human monoclonal antibodies that block the engagement of preS1 with NTCP and neutralize HBV and HDV with high potency. One antibody, 2H5-A14, functions at picomolar level and exhibited neutralization-activity-mediated prophylactic effects. It also acts therapeutically by eliciting antibody-Fc-dependent immunological effector functions that impose durable suppression of viral infection in HBV-infected mice, resulting in reductions in the levels of the small envelope antigen and viral DNA, with no emergence of escape mutants. Our results illustrate a novel antibody-Fc-dependent approach for HBV treatment and suggest 2H5-A14 as a novel clinical candidate for HBV prevention and treatment of chronic HBV infection.

Virus-like Particles Identify an HIV V1V2 Apex-Binding Neutralizing Antibody that Lacks a Protruding Loop.

  • Cale EM
  • Immunity
  • 2017 May 16

Literature context:


Most HIV-1-specific neutralizing antibodies isolated to date exhibit unusual characteristics that complicate their elicitation. Neutralizing antibodies that target the V1V2 apex of the HIV-1 envelope (Env) trimer feature unusually long protruding loops, which enable them to penetrate the HIV-1 glycan shield. As antibodies with loops of requisite length are created through uncommon recombination events, an alternative mode of apex binding has been sought. Here, we isolated a lineage of Env apex-directed neutralizing antibodies, N90-VRC38.01-11, by using virus-like particles and conformationally stabilized Env trimers as B cell probes. A crystal structure of N90-VRC38.01 with a scaffolded V1V2 revealed a binding mode involving side-chain-to-side-chain interactions that reduced the distance the antibody loop must traverse the glycan shield, thereby facilitating V1V2 binding via a non-protruding loop. The N90-VRC38 lineage thus identifies a solution for V1V2-apex binding that provides a more conventional B cell pathway for vaccine design.

Funding information:
  • Intramural NIH HHS - ZIA AI005023-15()
  • NIAID NIH HHS - R01 AI093278()
  • NIAID NIH HHS - R33 AI084714()

Tim29 is a novel subunit of the human TIM22 translocase and is involved in complex assembly and stability.

  • Kang Y
  • Elife
  • 2016 Aug 24

Literature context:


The TIM22 complex mediates the import of hydrophobic carrier proteins into the mitochondrial inner membrane. While the TIM22 machinery has been well characterised in yeast, the human complex remains poorly characterised. Here, we identify Tim29 (C19orf52) as a novel, metazoan-specific subunit of the human TIM22 complex. The protein is integrated into the mitochondrial inner membrane with it's C-terminus exposed to the intermembrane space. Tim29 is required for the stability of the TIM22 complex and functions in the assembly of hTim22. Furthermore, Tim29 contacts the Translocase of the Outer Mitochondrial Membrane, TOM complex, enabling a mechanism for transport of hydrophobic carrier substrates across the aqueous intermembrane space. Identification of Tim29 highlights the significance of analysing mitochondrial import systems across phylogenetic boundaries, which can reveal novel components and mechanisms in higher organisms.