Recombinant ifnα17 αi

Manufactured by PBL Assay Science

Recombinant IFNα17/αI is a laboratory reagent produced using recombinant DNA technology. It is a type I interferon protein that exhibits biological activity.

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Lab products found in correlation

Simoa assay, by Quanterix (3 mentions) Simoa assay kit, by Quanterix (1 mentions) Homebrew simoa assay, by Quanterix (1 mentions)

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IFNα17 (3 citations)

6 protocols using recombinant ifnα17 αi

Digital ELISA for Interferon Alpha Quantification

Cited 2 times

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Interferon alpha (IFN-α) protein levels were quantified with a digital-ELISA assay (Simoa, Quanterix) developed by Rodero et al.^{36 (link)}, in accordance with the instructions provided by the manufacturer of the Homebrew Simoa assay kit, with two autoantibodies specific for IFN-α isolated and cloned from two patients with autoimmune polyendocrinopathy syndrome type 1 (APSl)/autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED)^{50 (link)}. The 8H1 antibody clone was used to coat paramagnetic beads (0.3 mg/ml), as a capture antibody, and the 12H5 antibody was biotinylated (biotin-to-antibody ratio 30: 1) and used as the detector. SBG was used at concentrations of 0.3 μg/ml and 150 pM as a detector and revelation enzyme, respectively. Recombinant IFN-α17/-αI (PBL Assay Science) was used to generate a standard curve, after we had tested for cross-reactivity. Each sample was analyzed in duplicate and at a 1:3 dilution. The limit of detection (LOD) was calculated as the mean value +3SD (positivity with 99% confidence) of reactivity for all blank runs. The LOD, taking the dilution factor into account, was 0.19 fg/ml.

Kong X.F., Worley L., Rinchai D., Bondet V., Jithesh P.V., Goulet M., Nonnotte E., Rebillat A.S., Conte M., Mircher C., Gürtler N., Liu L., Migaud M., Elanbari M., Habib T., Ma C.S., Bustamante J., Abel L., Ravel A., Lyonnet S., Munnich A., Duffy D., Chaussabel D., Casanova J.L., Tangye S.G., Boisson-Dupuis S, & Puel A. (2020). Three copies of four interferon receptor genes underlie a mild type I interferonopathy in Down syndrome. Journal of clinical immunology, 40(6), 807-819.

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Ultrasensitive IFN-α Digital ELISA

Cited 2 times

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As previously reported (5 (link)), Simoa digital ELISA specific for IFN-α was developed using a Quanterix Homebrew Assay, and two autoantibodies specific for IFN-α were isolated and cloned from two APS1/APECED patients (59 (link)). The 8H1 antibody clone was used as a capture antibody after coating paramagnetic beads (0.3 mg/ml), and the 12H5 was biotinylated (biotin/Ab ratio = 30:1) and used as the detector. Recombinant IFN-α17/αI (PBL Assay Science) was used as a standard curve after cross-reactivity testing. The limit of detection was calculated by the mean value of all blank runs ± 3 SDs and was 0.23 fg/ml.

Smith N., Rodero M.P., Bekaddour N., Bondet V., Ruiz-Blanco Y.B., Harms M., Mayer B., Bader-Meunier B., Quartier P., Bodemer C., Baudouin V., Dieudonné Y., Kirchhoff F., Sanchez Garcia E., Charbit B., Leboulanger N., Jahrsdörfer B., Richard Y., Korganow A.S., Münch J., Nisole S., Duffy D, & Herbeuval J.P. (2019). Control of TLR7-mediated type I IFN signaling in pDCs through CXCR4 engagement—A new target for lupus treatment. Science Advances, 5(7), eaav9019.

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Ultrasensitive IFNα Quantification Assay

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Simoa IFNα assay was developed using a Quanterix Homebrew Simoa assay and two autoantibodies specific for IFNα isolated and cloned from two patients mutated in APS1 (causing autoimmune polyendocrinopathy with candidiasis and ectodermal dysplasia, APECED) patients as recently described16 (link),30 (link). The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg ml⁻¹), and the 12H5 was biotinylated (biotin/antibody ratio of 30:1) and used as the detector. Recombinant IFNα17/αI (PBL Assay Science) was used as a standard curve after cross-reactivity testing. The limits of detection were calculated by the mean value of all blank runs + 3 s.d. and was 0.23 fg ml⁻¹.

Dhir A., Dhir S., Borowski L.S., Jimenez L., Teitell M., Rötig A., Crow Y.J., Rice G.I., Duffy D., Tamby C., Nojima T., Munnich A., Schiff M., de Almeida C.R., Rehwinkel J., Dziembowski A., Szczesny R.J, & Proudfoot N.J. (2018). Mitochondrial double-stranded RNA triggers antiviral signalling in humans. Nature, 560(7717), 238-242.

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Quantitative Measurement of IFN-α Using Simoa

Cited 2 times

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As reported by Rodero et al.^{26 (link)}, a Simoa interferon alpha (IFN-α) assay was developed using a Quanterix Homebrew Simoa assay and two autoantibodies specific for IFN-α isolated and cloned from two APS1/APECED patients recently described^{24 (link)}. The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/mL), and the 12H5 was biotinylated (biotin/Ab ratio = 30/1) and used as the detector. Recombinant IFN-α17/αI (PBL Assay Science) was used as a standard curve after cross-reactivity testing. The limits of detection (LOD) were calculated by the mean value of all blank runs + 3SDs and was 0.23 fg/mL.

Rodero M.P., Tesser A., Bartok E., Rice G.I., Della Mina E., Depp M., Beitz B., Bondet V., Cagnard N., Duffy D., Dussiot M., Frémond M.L., Gattorno M., Guillem F., Kitabayashi N., Porcheray F., Rieux-Laucat F., Seabra L., Uggenti C., Volpi S., Zeef L.A., Alyanakian M.A., Beltrand J., Bianco A.M., Boddaert N., Brouzes C., Candon S., Caorsi R., Charbit M., Fabre M., Faletra F., Girard M., Harroche A., Hartmann E., Lasne D., Marcuzzi A., Neven B., Nitschke P., Pascreau T., Pastore S., Picard C., Picco P., Piscianz E., Polak M., Quartier P., Rabant M., Stocco G., Taddio A., Uettwiller F., Valencic E., Vozzi D., Hartmann G., Barchet W., Hermine O., Bader-Meunier B., Tommasini A, & Crow Y.J. (2017). Type I interferon-mediated autoinflammation due to DNase II deficiency. Nature Communications, 8, 2176.

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Quantitative IFNα Detection Assay

Cited 2 times

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As described by Rodero et al., the Simoa IFNα assay was developed using a Quanterix Homebrew Simoa assay according to the manufacturer’s instructions and utilising two autoantibodies specific for IFNα isolated and cloned from 2 APS1/APECED patients [9 (link), 12 (link)]. The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/mL), and the 12H5 was biotinylated (biotin/Ab ratio = 30/1) and used as the detector. Recombinant IFNα17/αI (PBL Assay Science) was used as a standard curve after cross-reactivity testing. The limits of detection (LOD) were calculated by the mean value of all blank runs + 3SDs and was 0.23 fg/ml.

Reynolds J.A., Briggs T.A., Rice G.I., Darmalinggam S., Bondet V., Bruce E., Khan M., Haque S., Chinoy H., Herrick A.L., McCarthy E.M., Zeef L., Hayes A., Duffy D., Parker B, & Bruce I.N. (2019). Type I interferon in patients with systemic autoimmune rheumatic disease is associated with haematological abnormalities and specific autoantibody profiles. Arthritis Research & Therapy, 21, 147.

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Ultrasensitive IFN-α Detection Assay

Cited 2 times

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The Simoa IFN-α assay (Rodero et al., 2017 (link)) was developed using a Quanterix Homebrew Simoa assay according to the manufacturer’s instructions, and using two autoantibodies specific for IFN-α isolated and cloned from two APS1/autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy patients as described (Meyer et al., 2016 (link); Rodero et al., 2017 (link)). The 8H1 antibody clone was used as a capture antibody after coating on paramagnetic beads (0.3 mg/ml), and the 12H5 was biotinylated (biotin/antibody ratio = 30/1) and used as the detector. Recombinant IFNα17/αI (PBL Assay Science) was used to produce a standard curve after cross-reactivity testing. The limit of detection was calculated as the mean value of all blank runs +3 SD and was 0.23 fg/ml.

Lepelley A., Martin-Niclós M.J., Le Bihan M., Marsh J.A., Uggenti C., Rice G.I., Bondet V., Duffy D., Hertzog J., Rehwinkel J., Amselem S., Boulisfane-El Khalifi S., Brennan M., Carter E., Chatenoud L., Chhun S., Coulomb l’Hermine A., Depp M., Legendre M., Mackenzie K.J., Marey J., McDougall C., McKenzie K.J., Molina T.J., Neven B., Seabra L., Thumerelle C., Wislez M., Nathan N., Manel N., Crow Y.J, & Frémond M.L. (2020). Mutations in COPA lead to abnormal trafficking of STING to the Golgi and interferon signaling. The Journal of Experimental Medicine, 217(11), e20200600.

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