Hydrophobic interaction chromatography used for purification of a Treponema Reiter ribonucleic acid antigen precipitating with antibodies in human syphilitic sera

Petersen, C S; Pedersen, Niels Tinggaard; Axelsen, N. H.

doi:10.1016/0003-2697(81)90716-8

Cited by 5 publications

(5 citation statements)

References 20 publications

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“…Hydrophobic interaction chromatography [6] has been successfully applied for the purification of a treponemal antigen [13]. In this study we obtained five fractions of A. fumigatus somatic antigen by step wise desorption of a Phenyl-Sepharose column.…”

Section: Discussionmentioning

confidence: 99%

Fractionation of <i>Aspergillus fumigatus</i> Antigens by Hydrophobic Interaction Chromatography and Gel Filtration

Schønheyder¹,

Andersen²

1984

Int Arch Allergy Immunol

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Aspergillus fumigatus somatic antigen prepared from young hyphae was fractionated by hydrophobic interaction chromatography followed by gel filtration. The antigenic activity of fractions was investigated by enzyme-linked immunosorbent assay, crossed immunoelectrophoresis, and immunodiffusion. By enzyme-linked immunosorbent assay, high levels of IgG antibodies were detected against an antigen fraction of approximate molecular weight 470,000 both in patients with aspergillosis (n = 3) and in healthy controls (n = 3). High levels of antibodies to four fractions of MW 250,000, 160,000, 160,000–50,000, and 50,000–25,000 were found only in patients. The MW 250,000 fraction exhibited catalase activity, and the MW 50,000–25,000 fraction had neutral protease activity. These purified aspergillus antigens may be useful in serodiagnosis of aspergillosis.

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Section: Discussionmentioning

confidence: 99%

Fractionation of <i>Aspergillus fumigatus</i> Antigens by Hydrophobic Interaction Chromatography and Gel Filtration

Schønheyder¹,

Andersen²

1984

Int Arch Allergy Immunol

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“…The purity of the antigen used in the present EL!SA is demonstrated, in Fig. I, and this antigen was shown to be RNA [9]. Compared with the flagellum ELISA and the sonicate ELISA, the RNA ELISA has a lower sensitivity both in primary syphilis (Group 1) and in treated syphilis (Group 4) (Table !).…”

Section: Discussionmentioning

confidence: 86%

“…RNA antifn'n. The RNA antigen was purified from T. pluigeilems hiotype Reiter as previoLisly described [9] by a simple one-step hydrophobic interaction chromatography method on phenyt-Sepharosc4B (Pharmacia Fine Chemicals, Uppsala. Sweden).…”

Section: Methodsmentioning

confidence: 99%

“…Antigen TR-d was purified from an ultrasonified Reiter treponeme culture by an immunoadsorbcnt technique using insolubilized IgG from syphilis patients [7]. This method had a low yield and recovery and was replaced by a rapid one-step hydrophobic interaction chromatography method [9] that made it possible to isolate 10 mg of pure antigen from 4 g wet weight of the Reiter treponeme culture. TR-d consisted of RNA withotit detectable protein, lipid.…”

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confidence: 99%

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Ribonucleic Acid Antigen from the Reiter Treponeme Used in an ELISA for Antibodies in Syphilis

1982

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Purified RNA from Treponema phagedenis biotype Reiter was used as antigen in an enzyme-linked immunosorbent assay (ELISA) for IgG antibodies in syphilis. The RNA ELISA was compared with the fluorescent treponemal antibody absorption test (FTA-ABS), the Treponema pallidum immobilization test (TPI), and with two ELISAs using purified flagellum from the Reiter treponeme as antigen (flagellum ELISA), and sonified Reiter treponeme culture as antigen (sonicate ELISA). A total of 729 sera from patients with and without syphilis were studied. The RNA ELISA had a lower sensitivity (P less than 0.01) in primary syphilis than the flagellum ELISA, the sonicate ELISA, and the FTA-ABS. In treated syphilis the RNA ELISA was also less sensitive than the TPI (P less than 0.01).

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“…TR-e has been isolated and identified as the periplasmatic flagellum or axial filament of the Reiter treponeme (11,17). TR-dl has been found to consist of RNA (12), and recently TR-c and TR-d2 have been purified and shown to be protein antigens (9; manuscript in preparation). TR-e has been shown to be very useful in enzyme-linked immunosorbent assays for the detection of immunoglobulin G (IgG) and IgM antibodies in syphilis (16,18).…”

mentioning

confidence: 99%

Purification of TR-b, a Reiter treponeme protein antigen precipitating with antibodies in human syphilitic sera

Petersen¹,

Pedersen²,

Axelsen³

1982

Infect Immun

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Reiter treponeme antigen, cross-reacting with an antigen in Treponema pallidum (Nichols pathogenic strain). Sera from patients with secondary syphilis contain precipitating antibodies against TR-b. The isolation of TR-b from a bacterial sonic extract is described here. It involved five fractionation steps: anion-exchange chromatography (DE-52 Whatman), gel filtration (Ac-A-22 Ultrogel), and affinity chromatography on phenyl-Sepharose CL 4B, iminodiacetic acid-Sepharose CL 4B, and lysine-Sepharose 4B, respectively. The purified TR-b was enriched 199 times compared with the starting material, and the recovery was 12%. TR-b was shown to be a protein; it did not bind to a series of lectins, and by gel filtration and polyacrylamide gel electrophoresis, the molecular weight was determined to be 610,000 to 630,000. It was found by sodium dodecyl sulfatepolyacrylamide gel electrophoresis to be composed of identical 70,000-dalton subunits. The isolated TR-b was immunologically pure when tested in crossed immunoelectrophoresis against polyspecific anti-Reiter immunoglobulin. The purified TR-b antigen was used for the production of a monospecific rabbit antiserum, giving strong fluorescence with both the Reiter treponeme and T. pallidum in an indirect immunofluorescence test.

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Hydrophobic interaction chromatography used for purification of a Treponema Reiter ribonucleic acid antigen precipitating with antibodies in human syphilitic sera

Cited by 5 publications

References 20 publications

Fractionation of <i>Aspergillus fumigatus</i> Antigens by Hydrophobic Interaction Chromatography and Gel Filtration

Fractionation of <i>Aspergillus fumigatus</i> Antigens by Hydrophobic Interaction Chromatography and Gel Filtration

Ribonucleic Acid Antigen from the Reiter Treponeme Used in an ELISA for Antibodies in Syphilis

Purification of TR-b, a Reiter treponeme protein antigen precipitating with antibodies in human syphilitic sera

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