Large scale recovery and purification of periplasmic recombinant protein from E. coli using expanded bed adsorption chromatography followed by new ion exchange media

Johansson, Hans J.; Jägersten, Christina; Shiloach, Joseph

doi:10.1016/0168-1656(96)01390-9

Cited by 71 publications

(32 citation statements)

References 7 publications

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“…Recombinant Pfs25 expressed in Pichia pastoris and rEPA from P. aeruginosa were prepared and characterized as described (25,26). OVA was from Sigma, St. Louis, MO.…”

Section: Methodsmentioning

confidence: 99%

Long-lasting and transmission-blocking activity of antibodies toPlasmodium falciparumelicited in mice by protein conjugates of Pfs25

Kübler-Kiełb

Majadly

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Malaria is a leading cause of morbidity and mortality, estimated to cause >1 million childhood deaths annually. Plasmodium falciparum causes the most severe form of the disease. There is as yet no licensed vaccine for this disease, despite over a half century of research. In this study, we investigated a transmission-blocking vaccine candidate, the ookinete surface protein Pfs25. Antibodies against Pfs25, drawn in during a bite, can block parasite development in the mosquito midgut, preventing transmission to other individuals. Pfs25 is a low-molecular-weight protein, by itself not immunogenic. To increase its immunogenicity, we investigated several methods of conjugating Pfs25 to itself and to other proteins: recombinant Pseudomonas aeruginosa exotoxin A, and ovalbumin, using amide, hydrazone, or thioether linkages. All conjugates were immunogenic and induced booster responses in mice. The scheme to form amide bonds between proteins by using adipic acid dihydrizide as a linker produced the most immunogenic conjugates. Adsorption of the conjugates onto aluminum hydroxide further increased the antibody response. Remarkably, the antibody levels 3 or 7 months after the last injection were significantly higher than those 1 wk after that injection. The observed transmission-blocking activity of immune sera correlated with antibody levels measured by ELISA. malaria ͉ vaccine O ne approach for a vaccine against malaria is to block transmission of the parasite from mosquitoes to humans. When ingested by a mosquito with the blood meal, antibodies against the sexual and the mosquito stage-specific surface antigens can block parasite development inside the vector (1). Four proteins have been identified as potential inducers of transmission-blocking antibodies (2-5). Two of these are expressed on the surface of gametes and in intracellular gameotocytes. The other two are the Plasmodium falciparum surface proteins with apparent molecular masses of 25 kDa (Pfs25) and 28 kDa (Pfs28), expressed exclusively on the zygote and ookinete surfaces during the mosquito stage of the infection. No antibody response to these two proteins has been shown in people infected with malaria and living in endemic countries (6). Pfs25 from different parts of the world has shown minimal variation in its amino acid sequence (7). This relative homogeneity, likely a result from not being subjected to immune pressure in the human host, makes Pfs25 an attractive candidate for a malaria transmission-blocking vaccine (8). Pfs25 is poorly immunogenic in mice and in humans, even if administered with adjuvant (9, 10). In this article, we show that Pfs25 bound onto itself or onto another protein induced high levels of transmission-blocking antibodies in mice.

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“…Recombinant Pfs25 expressed in Pichia pastoris and rEPA from P. aeruginosa were prepared and characterized as described (25,26). OVA was from Sigma, St. Louis, MO.…”

Section: Methodsmentioning

confidence: 99%

Long-lasting and transmission-blocking activity of antibodies toPlasmodium falciparumelicited in mice by protein conjugates of Pfs25

Kübler-Kiełb

Majadly

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…BSA (Sigma) was dialyzed against pyrogen-free water, sterile-filtered, and freeze-dried. Recombinant PA (rPA) from B. anthracis and recombinant exoprotein A (rEPA) from Pseudomonas aeruginosa were prepared and characterized (29,30).…”

Section: Methodsmentioning

confidence: 99%

Poly(γ-d-glutamic acid) protein conjugates induce IgG antibodies in mice to the capsule ofBacillus anthracis: A potential addition to the anthrax vaccine

Schneerson

Kübler-Kiełb

Liu

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

128

120

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“…EBA is an established procedure for the direct adsorption of proteins from crude feed stock, such as fermentation broth or biomass extract, eliminating the need for centrifugation and clarification, thereby greatly simplifying the recovery process (Anspach et al 1999;Chase 1994;Frej et al 1994;Johansson et al 1996;Shiloach and Kennedy 2000). In this operation, the adsorbent bed is expanded by an upward liquid flow which is controlled by a liquid distribution system.…”

Section: Introductionmentioning

confidence: 99%

Screen-less expanded bed column: new approach for the recovery and purification of a malaria transmission blocking vaccine candidate from Pichia pastoris

et al. 2006

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An experimental malaria transmission blocking vaccine antigen, Pfs25H, expressed and secreted from Pichia pastoris was recovered and purified using a screenless expanded bed column equipped with a rotating fluid distribution system. This column was able to accommodate feed stock, containing 30% biomass, at a flow rate of 300-400 cm/h without affecting column stability. This capability is three times higher than the capability of the expanded bed column currently in use, which is equipped with a perforated plate fluid distribution system; this design could accommodate biomass concentrations of only up to 10%. The screen-less design did not affect the binding capacity, purification level or process yield and, therefore, shorten the process. Purified Pfs25H of 6.4 g were recovered from 37 l of Pichia pastoris culture in one step.

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Large scale recovery and purification of periplasmic recombinant protein from E. coli using expanded bed adsorption chromatography followed by new ion exchange media

Cited by 71 publications

References 7 publications

Long-lasting and transmission-blocking activity of antibodies toPlasmodium falciparumelicited in mice by protein conjugates of Pfs25

Long-lasting and transmission-blocking activity of antibodies toPlasmodium falciparumelicited in mice by protein conjugates of Pfs25

Poly(γ-d-glutamic acid) protein conjugates induce IgG antibodies in mice to the capsule ofBacillus anthracis: A potential addition to the anthrax vaccine

Screen-less expanded bed column: new approach for the recovery and purification of a malaria transmission blocking vaccine candidate from Pichia pastoris

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