Purification of a recombinant protein expressed in yeast: optimization of analytical and preparative chromatography

Raymond, Frédérique; Rolland, Dominique; Gauthier, Marie; Jolivet, Michel

doi:10.1016/s0378-4347(97)00512-4

Cited by 21 publications

(15 citation statements)

References 19 publications

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“…If the PGA adsorption could occur at a pH lower than 4.8, probably higher PGA dynamic capacities would have been determined. That may also happens for the ionic adsorption of other enzymes as the dynamic capacities values reported in the literature for total protein adsorption (q Prot ) capacities are comparable with the adsorption capacities obtained in this work, with crude broths and using the same cation exchanger [14,[27][28][29].…”

Section: Eba Performance On the Adsorption Of Pga Onto Streamline Sp supporting

confidence: 88%

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Pinotti

Fonseca

Prazeres

et al. 2009

Biochemical Engineering Journal

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Section: Eba Performance On the Adsorption Of Pga Onto Streamline Sp supporting

confidence: 88%

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Pinotti

Fonseca

Prazeres

et al. 2009

Biochemical Engineering Journal

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“…GRA1 and GRA6 (kindly provided by M.-F. CesbronDelauw, Institut Pasteur, Lille, France) and GRA7 (4) (a gift from D. Jacobs and C. Dello, Immunogenetics, Ghent, Belgium) were expressed as fusion proteins with glutathione S-transferase (GST) in Escherichia coli. SAG1 was kindly provided by D. Rolland (Biomérieux) and produced in Schizosaccharomyces pombe yeast (15). Native GST or PBS served as negative controls.…”

Section: Methodsmentioning

confidence: 99%

Cellular Immune Responses to Recombinant Antigens in Pregnant Women Chronically Infected with Toxoplasma gondii

Fatoohi¹,

Cozon²,

Greenland

et al. 2002

Clin Vaccine Immunol

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The parasite Toxoplasma gondii can infect most mammals and birds, sometimes causing severe pathology. Primary infection during pregnancy can result in abortion or fetal defects. Host immunity, particularly cellular immunity towards antigenic peptides, can control infection, but an efficient vaccine is not yet available. We have evaluated T-cell responses to a crude soluble toxoplasma antigen (ST-Ag) and to five recombinant peptide antigens of cells in whole-blood cultures from 22 pregnant women with preexisting infections and from 7 pregnant negative controls. Cells from all infected patients but from none of the controls responded specifically to ST-Ag by expressing surface CD25 on culture. Responses to the recombinant antigens showed considerable variation between individuals. rGRA1 elicited a response in 16 of the 22 samples (73%), rSAG1 in 13, rGRA7 in 9, rGRA6-CT in 4, and rGRA6-NT in only 1. Most responding cells were CD4؉ . Cells from infected subjects cultured with ST-Ag all released high levels of gamma interferon (IFN-␥) into the culture supernatant (4,343 ؎ 2,536 pg/ml). Cells from 12 patients released IFN-␥ after culture with rGRA1 (130 ؎ 98 pg/ml), those from 10 patients released it after culture with rSAG1 (183 ؎ 128 pg/ml), and those from 4 patients released it after culture with rGRA7 (324 ؎ 374 pg/ml). Intensity of IFN-␥ production in response to the latter two recombinant antigens correlated with responses to ST-Ag (r ‫؍‬ 0.61 and 0.53, respectively; P < 0.01). Interleukin-4 was always absent from supernatants of cells stimulated with toxoplasma antigens. The heterogeneity of human responses to individual recombinant toxoplasma antigens should be considered in the design of potential vaccines.The protozoan parasite Toxoplasma gondii infests a wide range of mammalian hosts and some birds, usually with few pathological consequences. Human infestation is widespread and originates mainly, but not only, with cats. Incidence varies widely between geographical sites, but disease is rare, except in immunodepressed individuals and, especially, as a result of congenital infection. Cellular immunity provides an essential component of protection against T. gondii, and J. K. Frenkel (reviewed by Darcy and Santoro [2]) first demonstrated the key role played by T lymphocytes. Infection during pregnancy, especially during the third trimester, carries a high risk of mother-to-fetus transmission with severe consequences for the child. Immunocompetent mothers infected before pregnancy do not pass the parasite to their offspring in utero, even if reexposure to the parasite occurs during the critical period. The crucial T-cell responses to T. gondii may be measured (10), and we have recently determined that cytofluorimetric detection of CD25 expression after stimulation of whole-blood cultures with T. gondii antigen for 7 days is a sensitive and specific test, comparable to the 3 H-thymidine incorporation assay (5). The assay, which uses a crude soluble T. gondii antigen preparation (ST-Ag), was found to be useful ...

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“…The medium, culture and batch fermentation conditions have been previously described [33]. Additionally, four separate nitrogen sources were tested: (1) an acid digest of casein (HyCase SF, Sheffield Products, Norwich, NY, USA); (2) casein hydrochloric peptone (bio-Case, bioMérieux, Marcy l'Etoile, France); (3) casein tryptic peptone (bio-Trypcase, bioMérieux); and (4) yeast extract, yeast lysate (bioMérieux).…”

Section: Yeast Strain and Culture Processmentioning

confidence: 99%

Strategies for improving production and purification of a recombinant protein: rP30 of Toxoplasma gondii expressed in the yeast Schizosaccharomyces pombe

Rolland

Raymond

Gauthier

et al. 2008

Journal of Chromatography B

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Purification of a recombinant protein expressed in yeast: optimization of analytical and preparative chromatography

Cited by 21 publications

References 19 publications

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Cellular Immune Responses to Recombinant Antigens in Pregnant Women Chronically Infected with Toxoplasma gondii

Strategies for improving production and purification of a recombinant protein: rP30 of Toxoplasma gondii expressed in the yeast Schizosaccharomyces pombe

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