Pierce protein g spin plate

Manufactured by Thermo Fisher Scientific

The Pierce Protein G Spin Plate is a laboratory equipment used for the purification of antibodies from biological samples. It utilizes Protein G, a bacterial cell surface protein that binds to the Fc region of immunoglobulins, to capture and isolate antibodies. The spin plate format allows for efficient, small-scale antibody purification through a simple centrifugation process.

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Lab products found in correlation

Protein g elution buffer, by Thermo Fisher Scientific (2 mentions) Glycanassure data analysis software version 2, by Thermo Fisher Scientific (1 mentions) Glycanassure apts kit, by Thermo Fisher Scientific (1 mentions)

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Pierce™ Protein G Spin Plate for IgG Screening (2 citations)

7 protocols using pierce protein g spin plate

Purification and Epitope Profiling of IgG

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Polyclonal IgG were purified from plasma or sera from vaccinated, convalescent and naïve donors with a Pierce Protein G Spin Plate (ThermoFisher Scientific). Briefly, plasma and sera were diluted 1:4 in PBS before incubation with Protein G at room temperature for 30 min, with shaking at 600 rpm. Flowthrough was collected and incubated with the Protein G resin for an additional 15 min to ensure maximum binding. The protein G resin was washed four times with PBS before IgG was eluted with Protein G Elution Buffer (ThermoFisher Scientific). This buffer was neutralized with 1 M Tris pH 8.0 and buffer exchanged into PBS using a 40 kDa MWCO Zeba Plate. IgG was diluted to 100 μg/mL to assess epitope reactivity. Antibodies were added to a SAD200M chip (Carterra) with biotinylated peptide 154 as described above and binding was analyzed using the Epitope Software (Carterra).

Dacon C., Peng L., Lin T.H., Tucker C., Lee C.C., Cong Y., Wang L., Purser L., Cooper A.J., Williams J.K., Pyo C.W., Yuan M., Kosik I., Hu Z., Zhao M., Mohan D., Peterson M., Skinner J., Dixit S., Kollins E., Huzella L., Perry D., Byrum R., Lembirik S., Murphy M., Zhang Y., Yang E.S., Chen M., Leung K., Weinberg R.S., Pegu A., Geraghty D.E., Davidson E., Doranz B.J., Douagi I., Moir S., Yewdell J.W., Schmaljohn C., Crompton P.D., Mascola J.R., Holbrook M.R., Nemazee D., Wilson I.A, & Tan J. (2023). Rare, convergent antibodies targeting the stem helix broadly neutralize diverse betacoronaviruses. Cell Host & Microbe, 31(1), 97-111.e12.

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Purification of IgG from Plasma

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Bulk IgG was purified from plasma using Pierce Protein G Spin Plate (Thermo Fisher; catalog # 45204). IgG purity was confirmed by SDS gel.

Giron L.B., Dweep H., Yin X., Wang H., Damra M., Goldman A.R., Gorman N., Palmer C.S., Tang H.Y., Shaikh M.W., Forsyth C.B., Balk R.A., Zilberstein N.F., Liu Q., Kossenkov A., Keshavarzian A., Landay A, & Abdel-Mohsen M. (2021). Plasma Markers of Disrupted Gut Permeability in Severe COVID-19 Patients. Frontiers in Immunology, 12, 686240.

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Quantification of Plasma N-Glycans

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IgG was purified from 50 μl of plasma using the Pierce Protein G Spin Plate (Thermo Fisher catalog #45204). N-glycans were released using peptide-N-glycosidase F (PNGase F) and labeled with 8-aminopyrene-1,3,6-trisulfonic acid (APTS) using the GlycanAssure APTS Kit (Thermo Fisher, catalog #A33952), following the manufacturer’s protocol. The labeled N-glycans were analyzed using the 3500 Genetic Analyzer capillary electrophoresis system. The relative abundance of N-glycan structures was quantified by calculating the area under the curve of each glycan structure divided by the total glycans using the Applied Biosystems GlycanAssure Data Analysis Software Version 2.0.

Giron L.B., Liu Q., Adeniji O.S., Yin X., Kannan T., Ding J., Lu D.Y., Langan S., Zhang J., Azevedo J.L., Hanna D.B., Ofotokun I., Lazar J., Fischl M.A., Haberlen S., Macatangay B., Adimora A.A., Jamieson B.D., Rinaldo C., Merenstein D., Roan N.R., Kutsch O., Gange S., Wolinsky S., Witt M., Post W.S., Kossenkov A., Landay A., Frank I., Tien P.C., Gross R., Brown T.T, & Abdel-Mohsen M. (2023). Plasma Glycomic Markers of Accelerated Biological Aging During Chronic HIV Infection. bioRxiv.

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Protein G Purification of IgG

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Bulk IgG was purified from 50 µl plasma using Pierce™ Protein G Spin Plate (Thermo Fisher catalog# 45204). IgG purity was confirmed by SDS gel.

Giron L.B., Palmer C.S., Liu Q., Yin X., Papasavvas E., Sharaf R., Etemad B., Damra M., Goldman A.R., Tang H.Y., Johnston R., Mounzer K., Kostman J.R., Tebas P., Landay A., Montaner L.J., Jacobson J.M., Li J.Z, & Abdel-Mohsen M. (2021). Non-invasive plasma glycomic and metabolic biomarkers of post-treatment control of HIV. Nature Communications, 12, 3922.

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Polyclonal IgG Antibody Purification

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Polyclonal IgG antibodies from plasma or sera of vaccinated, convalescent, or naïve donors were purified using the Pierce Protein G Spin Plate (Thermo Scientific). Briefly, plasma or sera were diluted 1:4 in PBS and incubated with Protein G for 30 min, 600 rpm at room temperature. The flowthrough was collected and incubated with the Protein G resin for 15 min to ensure maximal binding. The Protein G resin was washed four times with PBS and the IgG was eluted with Protein G Elution Buffer (Thermo Scientific) and neutralized with 1 M Tris pH 8.0. Purified IgG was desalted using a 40 kDa MWCO Zeba Plate and diluted to 100 μg/mL to assess epitope reactivity.

Dacon C., Tucker C., Peng L., Lee C.C., Lin T.H., Yuan M., Cong Y., Wang L., Purser L., Williams J.K., Pyo C.W., Kosik I., Hu Z., Zhao M., Mohan D., Cooper A., Peterson M., Skinner J., Dixit S., Kollins E., Huzella L., Perry D., Byrum R., Lembirik S., Zhang Y., Yang E.S., Chen M., Leung K., Weinberg R.S., Pegu A., Geraghty D.E., Davidson E., Douagi I., Moir S., Yewdell J.W., Schmaljohn C., Crompton P.D., Holbrook M.R., Nemazee D., Mascola J.R., Wilson I.A, & Tan J. (2022). Broadly neutralizing antibodies target the coronavirus fusion peptide. bioRxiv.

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Protein G Purification of IgG

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Bulk IgG was purified from plasma using Pierce™ Protein G Spin Plate (Thermo Fisher).

GIRON L.B., PAPASAVVAS E., AZZONI L., YIN X., ANZUREZ A., DAMRA M., MOUNZER K., KOSTMAN J.R., SANNE I., FIRNHABER C.S., TATENO H., LIU Q., MONTANER L.J, & ABDEL-MOHSEN M. (2020). Plasma and Antibody Glycomic Biomarkers of Time to HIV Rebound and Viral Setpoint. AIDS (London, England), 34(5), 681-686.

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Purification and Analysis of IgG

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Bulk IgG was purified from plasma using Pierce™ Protein G Spin Plate (ThermoFisher Scientific, catalogue# 45204). IgG purity was confirmed by SDS gel. IgG subclasses were quantified by commercial ELISA kit (ThermoFisher Scientific, catalogue# 991000).

Giron L.B., Colomb F., Papasavvas E., Azzoni L., Yin X., Fair M., Anzurez A., Damra M., Mounzer K., Kostman J.R., Tebas P., O'Doherty U., Tateno H., Liu Q., Betts M.R., Montaner L.J, & Abdel-Mohsen M. (2020). Interferon-α alters host glycosylation machinery during treated HIV infection. EBioMedicine, 59, 102945.

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