Recombinant production of anti-HIV protein, griffithsin, by auto-induction in a fermentor culture

Giomarelli, Barbara; Schumacher, Kathryn M.; Taylor, Troy; Sowder, Raymond C.; Hartley, James L.; McMahon, James B.; Mori, Toshiyuki

doi:10.1016/j.pep.2005.10.014

Cited by 59 publications

(74 citation statements)

References 12 publications

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“…While both of these chimeric compounds would likely be highly effective microbicides, they have the disadvantage of being produced in a mammalian cell system (HEK cells), which would make it difficult to produce gram quantities of the inhibitor. In our system, Griff37 was produced from E. coli in shaker flasks and could likely be produced in large quantities by fermentation or in a plant-based expression system, both processes that were recently shown to be useful in making large quantities of functional griffithsin (22,41). Another effective HIV inhibitor was produced by linking two domains from CD4 with a single-chain variable region from antibody 17b, which is known to interact with gp120 on a site near its CCR5-binding region.…”

Section: Discussionmentioning

confidence: 99%

“…The amino acid sequence of the linker used for Griff37 as well as for C37CD4M33 C1F23 is SSSGG GGSGG GSSSG S, with minor variations between the different constructs due to cloning procedures. The gene for griffithsin and Griff37 includes a 6-histidine affinity tag as well as a thrombin cleavage site, similar to that previously described (22). The sequence for Griff37 is MGGSS HHHHH HSSGL VPRGS LTHRK FGGSG GSPFS GLSSI AVRSG SYLDA IIIDG VHHGG SGGNL SPTFT FGSGE YISNM TIRSG DYIDN ISFET NMGRR FGPYG GSGGS ANTLS NVKVI QINGS AGDYL DSLDI YYEQY SSSGG GGSGG GSSSG SHTTW MEWDR EINNY TSLIH SLIEE SQNQQ EKNEQ ELL.…”

Section: Methodsmentioning

confidence: 99%

“…This attempt was unsuccessful, so experiments proceeded in the presence of the His tag for griffithsin, Griff37, Griff37(Q652L), Griff37-shortlink, Griff37(I642D), and griffithsin covalently linked via a 16-amino-acid peptide linker with CD4M33 C1F23 (GriffCD4M33 C1F23 ). Others have shown that the presence of an N-terminal His tag does not affect the activity of griffithsin (22,40).…”

Section: Methodsmentioning

confidence: 99%

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Potent Strategy To Inhibit HIV-1 by Binding both gp120 and gp41

Kagiampakis

Gharibi

Mankowski

et al. 2011

Antimicrob Agents Chemother

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The development of an anti-HIV microbicide is critical in the fight against the spread of HIV. It is shown here that the covalent linking of compounds that bind gp120 with compounds that bind gp41 can inhibit HIV entry even more potently than individual inhibitors or noncovalent combinations. The most striking example involves griffithsin, a potent HIV inhibitor that binds to the surface of HIV gp120. While griffithsin inhibits HIV Env-mediated fusion in a CCR5-tropic cell-cell fusion assay with a 50% inhibitory concentration (IC 50 ) of 1.31 ؎ 0.87 nM and the gp41-binding peptide C37 shows an IC 50 of 18.2 ؎ 7.6 nM, the covalently linked combination of griffithsin with C37 (Griff37) has an IC 50 of 0.15 ؎ 0.05 nM, exhibiting a potency 8.7-fold greater than that of griffithsin alone. Similarly, in CXCR4-tropic cell-cell fusion assays, Griff37 is 5.2-fold more potent than griffithsin alone. In viral assays, both griffithsin and Griff37 inhibit HIV replication at midpicomolar levels, but the linked compound Griff37 is severalfold more potent than griffithsin alone against both CCR5-and CXCR4-tropic virus strains. Another example of this strategy is the covalently linked combination of peptide C37 with a variant of the gp120-binding peptide CD4M33 (L. Martin et al., Nat. Biotechnol. 21:71-76, 2003). Also, nuclear magnetic resonance (NMR) spectra for several of these compounds are shown, including, to our knowledge, the first published NMR spectrum for griffithsin.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Potent Strategy To Inhibit HIV-1 by Binding both gp120 and gp41

Kagiampakis

Gharibi

Mankowski

et al. 2011

Antimicrob Agents Chemother

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“…CV-N, SVN and GRFT used in in vitro studies were all produced in E. coli as previously reported [39][40][41]. GRFT used in animal studies was produced in Nicotiana benthamiana as previously reported [42].…”

Section: Reagentsmentioning

confidence: 99%

Antiviral lectins from red and blue-green algae show potent in vitro and in vivo activity against Hepatitis C virus

Takebe¹,

Saucedo²,

Lund³

et al. 2012

Planta Med

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Hepatitis C virus (HCV) infection is a significant public health problem with over 170,000,000 chronic carriers and infection rates increasing worldwide. Chronic HCV infection is one of the leading causes of hepatocellular carcinoma which was estimated to result in ,10,000 deaths in the United States in the year 2011. Current treatment options for HCV infection are limited to PEG-ylated interferon alpha (IFN-a), the nucleoside ribavirin and the recently approved HCV protease inhibitors telaprevir and boceprevir. Although showing significantly improved efficacy over the previous therapies, treatment with protease inhibitors has been shown to result in the rapid emergence of drug-resistant virus. Here we report the activity of two proteins, originally isolated from natural product extracts, which demonstrate low or sub-nanomolar in vitro activity against both genotype I and genotype II HCV. These proteins inhibit viral infectivity, binding to the HCV envelope glycoproteins E1 and E2 and block viral entry into human hepatocytes. In addition, we demonstrate that the most potent of these agents, the protein griffithsin, is readily bioavailable after subcutaneous injection and shows significant in vivo efficacy in reducing HCV viral titers in a mouse model system with engrafted human hepatocytes. These results indicate that HCV viral entry inhibitors can be an effective component of anti-HCV therapy and that these proteins should be studied further for their therapeutic potential.

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“…Prior work has suggested that GRFT can be cost effectively produced in tobacco plants [168,169]. Additionally, prior studies have suggested GRFT's weak immunogenicity and demonstrated low induction of inflammatory cytokines and chemokines in animal models [33,169].…”

Section: Griffithsin Structure and Mechanism Of Actionmentioning

confidence: 99%

Evaluation of the safety and pharmacokinetic profile of the broad spectrum antiviral lectin Griffithsin.

Barton¹,

Lynn²

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Recombinant production of anti-HIV protein, griffithsin, by auto-induction in a fermentor culture

Cited by 59 publications

References 12 publications

Potent Strategy To Inhibit HIV-1 by Binding both gp120 and gp41

Potent Strategy To Inhibit HIV-1 by Binding both gp120 and gp41

Antiviral lectins from red and blue-green algae show potent in vitro and in vivo activity against Hepatitis C virus

Evaluation of the safety and pharmacokinetic profile of the broad spectrum antiviral lectin Griffithsin.

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