Production of the 42-kDa fragment of Plasmodium falciparum merozoite surface protein 1, a leading malaria vaccine antigen, in Arabidopsis thaliana seeds

Lau, On Sun; Ng, David C.; Chan, Wendy Wing Lam; Chang, Sandra P.; Sun, Samuel S. M.

doi:10.1111/j.1467-7652.2010.00526.x

Cited by 21 publications

(13 citation statements)

References 49 publications

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“…For example, a maximum yield of 0.0035% TSP for PfMSP1 19 and 0.25% TSP for PyMSP4/5 were achieved in stably transformed tobacco plants (Ghosh et al 2002;Wang et al 2008). The maximum reported yield of Plasmodium antigens expressed by stable nuclear transformation was for PfMSP1 42 produced in Arabidopsis thaliana seeds, with a yield of up to 5% total protein (Lau et al 2010). Recent studies involving stable integration of transgenes into the chloroplast genome have also resulted in the accumulation of large quantities of foreign protein in various plant species, for example, fusion proteins of CTB-MSP1 and CTB-AMA1 were expressed at up to 13% and 10% TSP in tobacco and up to 7.3% and 6.1% TSP in lettuce, respectively (Davoodi-Semiromi et al 2009).…”

Section: Discussionmentioning

confidence: 99%

“…This result indicates the inherent difficulty of predicting expression, even for proteins with a similar structure, and suggests that empirical verification is required for any particular protein. Expression of PfMSP1 42 in A. thaliana has also demonstrated the importance of sequence optimisation and targeting strategy on protein accumulation (Lau et al 2010). By synthesizing a plant-optimised PfMSP1 42 cDNA and either targeting the recombinant protein to protein storage vacuoles or fusing it with a stable plant storage protein, Lau et al have achieved an impressive improvement in PfMSP1 42 expression, from an undetectable level to 5% of total extractable protein.…”

Section: Discussionmentioning

confidence: 99%

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Production, characterisation and immunogenicity of a plant-made Plasmodium antigen—the 19 kDa C-terminal fragment of Plasmodium yoelii merozoite surface protein 1

Chung

Wang

Webster

et al. 2011

Appl Microbiol Biotechnol

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Development of a safe, effective and affordable malaria vaccine is central to global disease control efforts. One of the most highly regarded proteins for inclusion in an asexual blood stage subunit vaccine is the 19-kDa C-terminal fragment of merozoite surface protein 1 (MSP1(19)). As production of vaccine antigens in plants can potentially overcome cost and delivery hurdles, we set out to produce MSP1(19) in plants, characterise the protein and test its immunogenicity using a mouse model. Plasmodium yoelii MSP1(19) (PyMSP1(19)) was produced in Nicotiana benthamiana using the MagnICON® deconstructed TMV-based viral vector. PyMSP1(19) yield of at least 23% total soluble protein (TSP;3-4 mg/g Fwt) were achieved using a codon-optimised construct that was targeted to the apoplast. Freeze-dried leaf powder contained at least 20 mg PyMSP1(19) per gram dry weight and the protein retained immunogenicity in this form for more than 2 years. Characterisation studies, including SDS-PAGE, mass spectrometry and circular dichroism, indicated that the plant-expressed PyMSP1(19) was similar to its Escherichia coli- and Saccharomyces cerevisiae-expressed counterparts. Purified plant-made PyMSP1(19) induced strong immune responses following intraperitoneal immunisation, although titres were lower than those induced by an equivalent dose of purified E. coli-expressed PyMSP1(19). The reason for this is uncertain but may be due to differences in the oligomerisation profile of the vaccines. The plant-made PyMSP1(19) vaccine was also found to be orally immunogenic when delivered alone or following immunisation with a PyMSP1(19) DNA vaccine. This study adds to an increasing body of research supporting the feasibility of plants as both a factory for the production of malaria antigens, and as a safe and affordable platform for oral delivery of a temperature-stable malaria vaccine.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Production, characterisation and immunogenicity of a plant-made Plasmodium antigen—the 19 kDa C-terminal fragment of Plasmodium yoelii merozoite surface protein 1

Chung

Wang

Webster

et al. 2011

Appl Microbiol Biotechnol

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“…An extensive array of expression platforms have been used to generate vaccine antigens against malaria, including synthetic peptides, viral delivery systems, bacteria, transgenic plants or animals, insect cells, mammalian cell lines and yeast [5]–[9]. The production of Plasmodium proteins for use in subunit vaccines using heterologous expression systems presents a number of challenges, since efficient expression of correctly folded proteins may be precluded by the inherent characteristics of many P. falciparum genes and their products, such as repetitive sequence content, large open reading frames, complex disulfide bonding patterns and the high AT content of P. falciparum DNA [10], [11].…”

Section: Introductionmentioning

confidence: 99%

A Novel Malaria Vaccine Candidate Antigen Expressed in Tetrahymena thermophila

Cowan

Bockau²,

Eleni-Muus³

et al. 2014

PLoS ONE

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Development of effective malaria vaccines is hampered by the problem of producing correctly folded Plasmodium proteins for use as vaccine components. We have investigated the use of a novel ciliate expression system, Tetrahymena thermophila, as a P. falciparum vaccine antigen platform. A synthetic vaccine antigen composed of N-terminal and C-terminal regions of merozoite surface protein-1 (MSP-1) was expressed in Tetrahymena thermophila. The recombinant antigen was secreted into the culture medium and purified by monoclonal antibody (mAb) affinity chromatography. The vaccine was immunogenic in MF1 mice, eliciting high antibody titers against both N- and C-terminal components. Sera from immunized animals reacted strongly with P. falciparum parasites from three antigenically different strains by immunofluorescence assays, confirming that the antibodies produced are able to recognize parasite antigens in their native form. Epitope mapping of serum reactivity with a peptide library derived from all three MSP-1 Block 2 serotypes confirmed that the MSP-1 Block 2 hybrid component of the vaccine had effectively targeted all three serotypes of this polymorphic region of MSP-1. This study has successfully demonstrated the use of Tetrahymena thermophila as a recombinant protein expression platform for the production of malaria vaccine antigens.

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“…Turpen et al (1995) were the first to report the expression of a malaria antigen in the tobacco plant. Recently published works indicate the expression of antigens against malaria in lettuce and tobacco (Davoodi-semiromi et al, 2010), Arabidopsis thaliana seeds (Lau et al, 2010), and green algae (Dauvillée et al, 2010). The malaria antigens used were AMA1, MSP1 (Dauvillee et al, 2010;Davoodi-semiromi et al, 2010), and MSP1 42 (Lau et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Transgenic tomato plants expressing the antigen gene PfCP-2.9 of Plasmodium falciparum

Kantor

Sestraş

Chowdhury

2013

Pesq. agropec. bras.

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-The objective of this work was to obtain transgenic tomato plants expressing the PfCP-2.9 protein (a chimera of the antigens MSP1 and AMA1 of Plasmodium falciparum). Cotyledons of seven-day-old tomatoes, cultivar Summers, were transformed via Agrobacterium tumefaciens. Transgenic expression in the T0 plants was verified in the DNA extracted from fruits. PCR analysis was used to test the presence of the gene of interest in the T1 generation. Reverse transcriptase PCR provided evidence of gene expression at the RNA level, and Western blot analysis confirmed the presence of the protein of interest in the T1 plants. This is the first report of successful transformation with the expression of a malaria antigen (PfCP-2.9) in transgenic tomato plants from the T0 and T1 generations.Index terms: gene expression, genetic transformation, malaria, plant-made vaccines. Plantas transgênicas de tomate expressando o gene do antígeno PfCP-2.9 de Plasmodium falciparumResumo -O objetivo deste trabalho foi obter plantas transgênicas de tomate que expressem a proteína PfCP-2.9 (uma quimera dos antígenos MSP1 e AMA1 de Plasmodium falciparum). Cotilédones de tomate, cultivar Summers, com sete dias de idade, foram transformados via Agrobacterium tumefaciens. A expressão transgênica nas plantas T0 foi verificada no DNA extraído dos frutos. A análise por PCR foi utilizada para testar a presença do gene de interesse na geração T1. A evidência da expressão do gene no RNA foi constatada por meio da PCR de transcriptase reversa, e a análise "Western blot" confirmou a presença da proteína de interesse nas plantas T1. Este é o primeiro relato de transformação bem sucedida com a expressão de um antígeno da malária (PfCP-2,9) em plantas transgênicas de tomate da geração T0 e T1.Termos para indexação: expressão gênica, transformação genética, malária, vacina a partir de plantas.

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Production of the 42-kDa fragment of Plasmodium falciparum merozoite surface protein 1, a leading malaria vaccine antigen, in Arabidopsis thaliana seeds

Cited by 21 publications

References 49 publications

Production, characterisation and immunogenicity of a plant-made Plasmodium antigen—the 19 kDa C-terminal fragment of Plasmodium yoelii merozoite surface protein 1

Production, characterisation and immunogenicity of a plant-made Plasmodium antigen—the 19 kDa C-terminal fragment of Plasmodium yoelii merozoite surface protein 1

A Novel Malaria Vaccine Candidate Antigen Expressed in Tetrahymena thermophila

Transgenic tomato plants expressing the antigen gene PfCP-2.9 of Plasmodium falciparum

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