Plant Molecular Farming
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Norway to get world's last dose of ZMapp

Norway to get world's last dose of ZMapp | Plant Molecular Farming | Scoop.it

The Norwegian woman, infected by the Ebola in Sierra Leone and currently receiving treatment in Oslo, will get the last dose of the virus treatment medicine ZMapp

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...and yet again, the emphasis is on how slow it is to make it - when the whole point of biofarming and transient expression is that it is supposed to be QUICK to make things, and easy to scale up production!!

What is the problem here?  

...and Kenneth Palmer told me just today (Friday 10th Oct) what it is:
"You may not be aware that the human dose of Zmapp is 12 grams per patient, 3 infusions of 4 grams each.  Check the dose in recent Nature paper. If yield of one antibody is 100 mg per kg and you have to produce three antibodies for Zmapp... If you do the arithmetic you will see why the process is "slow"".

So.... Doing just that, you end up with 30 kg N benthamiana per gm of ZMapp as a best-case yield - meaning 360 kg PER PATIENT.

That's a LOT of N benth - and tooling up for that sort of plant production takes time. Thanks, Kenneth!

I would be VERY interested in a cost breakdown of ZMapp vs CHO cell-produced MAbs - because producing at that sort of scale MUST be prohibitively expensive in stainless steel?

 

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Plant Molecular Farming
A news site for the activities of members of the International Society for Plant Molecular Farming, as well as for papers of interest in the field
Curated by Ed Rybicki
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Plants Against Corona

Plants Against Corona | Plant Molecular Farming | Scoop.it
The logo of an alliance of plant biotechnology companies making products derived from SARS-CoV-2. Global partners in the network include Canadian company Plantform and United Kingdom-based Leaf Expressions, both of whom will assist with scaling up production of the antigens to meet global demands. Academic partners include Plantvax (from the United States) and the Department of Applied Genetics and 3 Cell Biology (DAGZ) in the University of Natural Resources and Life Sciences, Vienna. They will be using the genetic constructs for their own research and development to further expand the community. Quebec-based vertical farming equipment design company, Inno-3B, aims to assist the Plants Against Corona initiative in rapidly scaling up their plantgrowth capacity.
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Molecular Farming Means the Next Vaccine Could be Edible and Grown in a Plant - SoylentNews

Molecular Farming Means the Next Vaccine Could be Edible and Grown in a Plant -- article related to Science and The Main Page.
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Protection of Newborn Macaques by Plant-Derived HIV Broadly Neutralizing Antibodies: a Model for Passive Immunotherapy during Breastfeeding

Protection of Newborn Macaques by Plant-Derived HIV Broadly Neutralizing Antibodies: a Model for Passive Immunotherapy during Breastfeeding | Plant Molecular Farming | Scoop.it
Preventing human immunodeficiency virus (HIV) infection in newborns by vertical transmission remains an important unmet medical need in resource-poor areas where antiretroviral therapy (ART) is not available and mothers and infants cannot be treated prepartum or during the breastfeeding period. In the present study, the protective efficacy of the potent HIV-neutralizing antibodies PGT121 and VRC07-523, both produced in plants, were assessed in a multiple-SHIV (simian-human immunodeficiency virus)-challenge breastfeeding macaque model. Newborn macaques received either six weekly subcutaneous injections with PGT121 alone or as a cocktail of PGT121-LS plus VRC07-523-LS injected three times every 2 weeks. Viral challenge with SHIVSF162P3 was twice weekly over 5.5 weeks using 11 exposures. Despite the transient presence of plasma viral RNA either immediately after the first challenge or as single-point blips, the antibodies prevented a productive infection in all babies with no sustained plasma viremia, compared to viral loads ranging from 103 to 5 × 108 virions/ml in four untreated controls. No virus was detected in peripheral blood mononuclear cells (PBMCs), and only 3 of 159 tissue samples were weakly positive in the treated babies. Newborn macaques proved to be immunocompetent, producing transient anti-Env antibodies and anti-drug antibody (ADA), which were maintained in the circulation after passive broadly neutralizing antibody clearance. ADA responses were directed to the IgG1 Fc CH2-CH3 domains, which has not been observed to date in adult monkeys passively treated with PGT121 or VRC01. In addition, high levels of VRC07-523 anti-idiotypic antibodies in the circulation of one newborn was concomitant with the rapid elimination of VRC07. Plant-expressed antibodies show promise as passive immunoprophylaxis in a breastfeeding model in newborns. IMPORTANCE Plant-produced human neutralizing antibody prophylaxis is highly effective in preventing infection in newborn monkeys during repeated oral exposure, modeling virus in breastmilk, and offers advantages in cost of production and safety. These findings raise the possibility that anti-Env antibodies may contribute to the control of viral replication in this newborn model and that the observed immune responsiveness may be driven by the long-lived presence of immune complexes.
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Your next vaccine could be grown in a tobacco plant

Your next vaccine could be grown in a tobacco plant | Plant Molecular Farming | Scoop.it
Long-awaited plant-based vaccine technology could help COVID-19 shots reach developing countries.
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I know, I know; I'm biased...B-)
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Metabolic engineering of rice endosperm towards higher vitamin B1 accumulation

Rice is a major food crop to approximately half of the human population. Unfortunately, the starchy endosperm, which is the remaining portion of the seed after polishing, contains limited amounts of micronutrients. Here, it is shown that this is particularly the case for thiamin (vitamin B1). Therefore, a tissue-specific metabolic engineering approach was conducted, aimed at enhancing the level of thiamin specifically in the endosperm. To achieve this, three major thiamin biosynthesis genes, THIC, THI1 and TH1, controlled by strong endosperm-specific promoters, were employed to obtain engineered rice lines. The metabolic engineering approaches included ectopic expression of THIC alone, in combination with THI1 (bigenic) or combined with both THI1 and TH1 (trigenic). Determination of thiamin and thiamin biosynthesis intermediates reveals the impact of the engineering approaches on endosperm thiamin biosynthesis. The results show an increase of thiamin in polished rice up to threefold compared to WT, and stable upon cooking. These findings confirm the potential of metabolic engineering to enhance de novo thiamin biosynthesis in rice endosperm tissue and aid in steering future biofortification endeavours.
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Plant‐based production can result in covalent cross‐linking of proteins

Plant‐based production can result in covalent cross‐linking of proteins | Plant Molecular Farming | Scoop.it

Antibodies, antigens and enzymes for replacement therapies and virus-like particles (VLPs) have all been produced successfully in plants as part of the concept of ‘Molecular Farming’ (Lomonossoff and D’Aoust, 2016). There have been several differences noted between plant-expressed proteins, and their equivalents produced in other systems, such as CHO cells, particularly regarding their glycosylation. However, several publications have also indicated that preparations of plant-expressed proteins, including antibodies, VLPs and soluble molecules such as HIV gp120, have a higher proportion of multimers or aggregates than their CHO- or yeast-expressed equivalents (e.g. Mechtcheriakova et al., 2006; Ramessar et al., 2008; Rosenberg et al., 2013). The cause of this has not been investigated in detail because the molecules expressed in the different systems are often not identical and may have been purified to different extents.

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A new bromelain-based enzyme for the release of Dupuytren’s contracture 

A new bromelain-based enzyme for the release of Dupuytren’s contracture  | Plant Molecular Farming | Scoop.it
ObjectivesInjectable Bromelain Solution (IBS) is a modified investigational derivate of the medical grade bromelain-debriding pharmaceutical agent (NexoBrid) studied and approved for a rapid (four-...
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The key role of terminators on the expression and post‐transcriptional gene silencing of transgenes

The key role of terminators on the expression and post‐transcriptional gene silencing of transgenes | Plant Molecular Farming | Scoop.it

Transgenes have become essential to modern biology, being an important tool in functional genomic studies and also in the development of biotechnological products. One of the major challenges in the generation of transgenic lines concerns the expression of transgenes, which, compared to endogenes, are particularly susceptible to silencing mediated by small RNAs (sRNAs). Several reasons have been put forward to explain why transgenes often trigger production of sRNAs, such as the high level of expression induced by commonly‐used strong constitutive promoters, the lack of introns and features resembling virus and other exogenous sequences. However, the relative contributions of the different genomic elements in protecting genes from the silencing machinery and their molecular mechanisms remain unclear. Here, we present the results of a mutagenesis screening conceived to identify features involved in the protection of endogenes against becoming template for the production of sRNAs. Interestingly, all of the recovered mutants had alterations in genes with proposed function in transcription termination, suggesting a central role of terminators in this process. Indeed, using a GFP reporter system, we show that among different genetic elements tested, the terminator sequence had the greatest effect on transgene‐derived sRNA accumulation and that a well‐defined poly(A) site might be especially important. Finally, we describe an unexpected mechanism, where transgenes containing certain intron/terminator combinations lead to an increase in the production of sRNAs, which seems to interfere with splicing.

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‘Alarming’ use of critical human antibiotics on crops

Global survey finds ‘extensive’ use of antibiotics in crop production.
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Palliative Canadians endure punishing waits while Health Minister delays approval of Psilocybin mushrooms

Palliative Canadians endure punishing waits while Health Minister delays approval of Psilocybin mushrooms | Plant Molecular Farming | Scoop.it
VICTORIA, British Columbia, June 17, 2020 (GLOBE NEWSWIRE) -- Pressure grows on Health Minister, Patty Hajdu, as palliative Canadians wait for the health...
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GM risk assessments: the importance of in planta studies in the sustainable management of GM plants

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Italy and Israel bet on GM microalgae to develop edible COVID vaccine

Italy and Israel bet on GM microalgae to develop edible COVID vaccine | Plant Molecular Farming | Scoop.it
The rush to develop a vaccine for COVID-19 has extended to Italy and Israel, where scientists are using the tools of genetic engineering to develop algae-based edible vaccines against the disease caused by the novel coronavirus.
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Shotguns vs Lasers: Identifying barriers and facilitators to scaling-up plant molecular farming for high-value health products

Shotguns vs Lasers: Identifying barriers and facilitators to scaling-up plant molecular farming for high-value health products | Plant Molecular Farming | Scoop.it
Plant molecular farming (PMF) is a convenient and cost-effective way to produce high-value recombinant proteins that can be used in the production of a range of health products, from pharmaceutical therapeutics to cosmetic products. New plant breeding techniques (NPBTs) provide a means to enhance PMF systems more quickly and with greater precision than ever before. However, the feasibility, regulatory standing and social acceptability of both PMF and NPBTs are in question. This paper explores the perceptions of key stakeholders on two European Union (EU) Horizon 2020 programmes–Pharma-Factory and Newcotiana–towards the barriers and facilitators of PMF and NPBTs in Europe. One-on-one qualitative interviews were undertaken with N = 20 individuals involved in one or both of the two projects at 16 institutions in seven countries (Belgium, France, Germany, Italy, Israel, Spain and the UK). The findings indicate that the current EU regulatory environment and the perception of the public towards biotechnology are seen as the main barriers to scaling-up PMF and NPBTs. Competition from existing systems and the lack of plant-specific regulations likewise present challenges for PMF developing beyond its current niche. However, respondents felt that the communication of the benefits and purpose of NPBT PMF could provide a platform for improving the social acceptance of genetic modification. The importance of the media in this process was highlighted. This article also uses the multi-level perspective to explore the ways in which NPBTs are being legitimated by interested parties and the systemic factors that have shaped and are continuing to shape the development of PMF in Europe.
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Oral MucoRice-CTB vaccine for safety and microbiota-dependent immunogenicity in humans: a phase 1 randomised trial

Oral MucoRice-CTB vaccine for safety and microbiota-dependent immunogenicity in humans: a phase 1 randomised trial | Plant Molecular Farming | Scoop.it
Participants given MucoRice-CTB showed increased CTB-specific serum IgG and IgA antibody
concentrations without inducing serious adverse events, indicating that MucoRice-CTB
could be a safe and potent vaccine to prevent diarrhoeal disease. MucoRice-CTB induced
neutralising antibodies against diarrhoeal toxins in a gut microbiota-dependent manner.
A similar phase 1 trial will be done with participants of other ethnicities to substantiate
our findings.
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I don't know why a bigger deal is not being made of this: mainstreaming plant-made vaccines, and for bacteria this time!
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Study Shows High-Flavonoid Foods Protect Against Cognitive Decline

Study Shows High-Flavonoid Foods Protect Against Cognitive Decline | Plant Molecular Farming | Scoop.it
Cognitive decline is a common condition from many millions of people suffer. As we age, our bodies go through a myriad of physical and mental changes. One such age-related mental change is cognitive decline. It involves a decline in mental awareness and aptitude.
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Enhanced Ability of Plant-Derived PGT121 Glycovariants To Eliminate HIV-1-Infected Cells

Enhanced Ability of Plant-Derived PGT121 Glycovariants To Eliminate HIV-1-Infected Cells | Plant Molecular Farming | Scoop.it
The activity of broadly neutralizing antibodies (bNAbs) targeting HIV-1 depends on pleiotropic functions, including viral neutralization and the elimination of HIV-1-infected cells. Several in vivo studies have suggested that passive administration of bNAbs represents a valuable strategy for the prevention or treatment of HIV-1. In addition, different strategies are currently being tested to scale up the production of bNAbs to obtain the large quantities of antibodies required for clinical trials. Production of antibodies in plants permits low-cost and large-scale production of valuable therapeutics; furthermore, pertinent to this work, it also includes an advanced glycoengineering platform. In this study, we used Nicotiana benthamiana to produce different Fc-glycovariants of a potent bNAb, PGT121, with near-homogeneous profiles and evaluated their antiviral activities. Structural analyses identified a close similarity in overall structure and glycosylation patterns of Fc regions for these plant-derived Abs and mammalian cell-derived Abs. When tested for Fc-effector activities, afucosylated PGT121 showed significantly enhanced FcγRIIIa interaction and antibody dependent cellular cytotoxicity (ADCC) against primary HIV-1-infected cells, both in vitro and ex vivo. However, the overall galactosylation profiles of plant PGT121 did not affect ADCC activities against infected primary CD4+ T cells. Our results suggest that the abrogation of the Fc N-linked glycan fucosylation of PGT121 is a worthwhile strategy to boost its Fc-effector functionality. IMPORTANCE PGT121 is a highly potent bNAb and its antiviral activities for HIV-1 prevention and therapy are currently being evaluated in clinical trials. The importance of its Fc-effector functions in clearing HIV-1-infected cells is also under investigation. Our results highlight enhanced Fc-effector activities of afucosylated PGT121 MAbs that could be important in a therapeutic context to accelerate infected cell clearance and slow disease progression. Future studies to evaluate the potential of plant-produced afucosylated PGT121 in controlling HIV-1 replication in vivo are warranted.
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Plant-derived vaccines

Plant-derived vaccines | Plant Molecular Farming | Scoop.it
Quebec-based Medicago is transforming the use of plant-based technologies to rapidly develop and produce novel vaccines and therapeutic proteins.
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Expert opinions on the regulation of plant genome editing 

Expert opinions on the regulation of plant genome editing  | Plant Molecular Farming | Scoop.it

Global food security is largely affected by factors such as environmental (e.g. drought, flooding), social (e.g. gender inequality), socio-economic (e.g. overpopulation, poverty) and health (e.g. diseases). In response, extensive public and private investment in agricultural research has focused on increasing yields of staple food crops and developing new traits for crop improvement. New breeding techniques pioneered by genome editing have gained substantial traction within the last decade, revolutionizing the plant breeding field. Both industry and academia have been investing and working to optimize the potentials of gene editing and to bring derived crops to market. The spectrum of cutting-edge genome editing tools along with their technical differences has led to a growing international regulatory, ethical and societal divide. This article is a summary of a multi-year survey project exploring how experts view the risks of new breeding techniques, including genome editing and their related regulatory requirements. Surveyed experts opine that emerging biotechnologies offer great promise to address social and climate challenges, yet they admit that the market growth of genome-edited crops will be limited by an ambiguous regulatory environment shaped by societal uncertainty.

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Enhanced vitamin E content in an Indica rice cultivar harbouring two transgenes from Arabidopsis thaliana involved in tocopherol biosynthesis pathway

Enhanced vitamin E content in an Indica rice cultivar harbouring two transgenes from Arabidopsis thaliana involved in tocopherol biosynthesis pathway | Plant Molecular Farming | Scoop.it
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The α-tubulin of Laodelphax striatellus mediates the passage of rice stripe virus (RSV) and enhances horizontal transmission

The α-tubulin of Laodelphax striatellus mediates the passage of rice stripe virus (RSV) and enhances horizontal transmission | Plant Molecular Farming | Scoop.it
Author summary Over 70% of all known plant viruses are transmitted by specific arthropods, mainly including planthoppers, leafhoppers, aphids and whiteflies. Plant viruses with persistent relationships must overcome multiple barriers.
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Heterologous expression of biologically active Mambalgin-1 peptide as a new potential anticancer, using a PVX-based viral vector in Nicotiana benthamiana

Heterologous expression of biologically active Mambalgin-1 peptide as a new potential anticancer, using a PVX-based viral vector in Nicotiana benthamiana | Plant Molecular Farming | Scoop.it
Mambalgin-1 is a peptide that acts as a potent analgesic through inhibiting acid-sensing ion channels (ASIC) in nerve cells. Research has shown that ASIC channels are involved in the proliferation and growth of cancer cells; therefore, Mambalgin-1 can be a potential anti-cancer by inhibiting these channels. In the present study, the Nicotiana benthamiana codon optimized Mambalgin-1 gene was synthesized and cloned in PVX (potato virus X) viral vector. The two cultures of Agrobacterium containing Mambalgin-1 and P19 silencing suppressor genes were co-agroinfiltrated into N. benthamiana leaves. Five days post infiltration, the production of recombinant Mambalgin-1 was determined by western blotting. For biological activity, MTT (3(4, 5-dimethylthiazole-2-yl)-2, 5-diphenyltetrazolium bromide) was analyzed for the cytotoxicity recombinant Mambalgin-1 from the transformed plants on nervous (SH-SY5Y) and breast (MCF7) cancer cells. The results showed that the plants expressing open reading frame of Mambalgin-1 showed recombinant 7.4 kDa proteins in the entire plant extract. In the MTT test, it was found that Mambalgin-1 had cytotoxic effects on SH-SY5Y cancer cells, yet no effects on MCF7 cancer cells were observed. According to the results, the expression of the biologically active recombinant Mambalgin-1 in the transformed plant leaves was confirmed and Mambalgin-1 can also have anti-cancer (inhibition of ASIC channels) potential along with its already known analgesic effect. The Mambalgin-1 transiently expressed in Nicotiana benthamiana plant and its anti-cancer effect on SH-SY5Y nerve cancer cells, since ASIC channels are involved in the proliferation of cancer cells, was confirmed.
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Plant Gene Editing Through De Novo Induction of Meristems

Plant gene editing is typically performed by delivering reagents such as Cas9 and single guide RNAs to explants in culture. Edited cells are then induced to differentiate into whole plants by exposure to various hormones. The creation of edited plants through tissue culture is often inefficient, tim …
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Test the COVID Vax for Glyphosate! WARNING: NONSENSE ALERT!

Test the COVID Vax for Glyphosate! WARNING: NONSENSE ALERT! | Plant Molecular Farming | Scoop.it
Moms Across America - From Seed to Supermarket We're Forging a New Standard for a Healthy Planet...
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Calreticulin co-expression supports high level production of a recombinant SARS-CoV-2 spike mimetic in Nicotiana benthamiana | bioRxiv

An effective prophylactic vaccine is urgently needed to protect against SARS-CoV-2 infection. The viral spike, which mediates entry into cells by interacting with the host angiotensin-converting enzyme 2, is the primary target of most vaccines in development. These vaccines aim to elicit protective immunity against the glycoprotein by use of inactivated virus, vector-mediated delivery of the antigen in vivo, or by direct immunization with the purified antigen following expression in a heterologous system. These approaches are mostly dependent on the growth of mammalian or insect cells, which requires a sophisticated infrastructure that is not generally available in developing countries due to the incumbent costs which are prohibitive. Plant-based subunit vaccine production has long been considered as a cheaper alternative, although low expression yields and differences along the secretory pathway to mammalian cells have posed a challenge to producing certain viral glycoproteins. Recent advances that have enabled many of these constraints to be addressed include expressing the requisite human proteins in plants to support the maturation of the protein of interest. In this study we investigated these approaches to support the production of a soluble and putatively trimeric SARS-CoV-2 spike mimetic in Nicotiana benthamiana via transient Agrobacterium-mediated expression. The co-expression of human calreticulin dramatically improved the accumulation of the viral spike, which was barely detectable in the absence of the co-expressed accessory protein. The viral antigen was efficiently processed even in the absence of co-expressed furin, suggesting that processing may have occurred at the secondary cleavage site and was mediated by an endogenous plant protease. In contrast, the spike was not efficiently processed when expressed in mammalian cells as a control, although the co-expression of furin improved processing considerably. This study demonstrates the feasibility of molecular engineering to improve the production of viral glycoproteins in plants, and supports plant-based production of SARS-CoV-2 spike-based vaccines and reagents for serological assays.

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GMO tomato as edible COVID vaccine? Mexican scientists work to make it a reality

GMO tomato as edible COVID vaccine? Mexican scientists work to make it a reality | Plant Molecular Farming | Scoop.it
While large companies and public sector consortiums in the United States, Canada, China and Europe are running at full speed to develop a vaccine grown in genetically modified (GM) tobacco plants, a research group at a Mexican university is working toward the same objective, but with a different and innovative strategy. They are using bioinformatics …
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Assessments of hepatitis B virus-like particles and Crm197 as carrier proteins in melioidosis glycoconjugate vaccines | bioRxiv

The Tier 1 select agent Burkholderia pseudomallei is the causative agent of melioidosis, a global pathogen and a major cause of pneumonia and sepsis for which no licensed vaccines currently exist. Previous work has shown the potential for Burkholderia capsular polysaccharide (CPS) to be used as a vaccine antigen but the T-cell independent nature of the immune response to this molecule requires the use of this polysaccharide as a glycoconjugate for vaccination. Recent studies have focussed on the use of Crm197 (a non-toxic mutant protein derived from diphtheria toxin) as the carrier but there are concerns regarding its potential to cause interference with other vaccines containing Crm197. Therefore research with alternative carrier proteins would be beneficial. In this study, CPS was isolated from the non-pathogenic B. thailandensis strain E555. This was chemically conjugated to Crm197, or Tandem Core™ virus-like particles (TCVLP) consisting of hepatitis B core protein, which is the first documented use of VLPs in melioidosis vaccine development. Analysis of CPS-specific IgG antibody titres showed that mice vaccinated with the Crm197 conjugate generated significantly higher titres than the mice that received TCVLP-CPS but both conjugate vaccines were able to protect mice against intraperitoneal B. pseudomallei strain K96243 challenges of multiple median lethal doses.

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This is potentially a big deal: being able to make REALLY cheap carriers for glycoconjugate vaccines could revolutionise their application.
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