Viral entry experiments were carried out as previously described.49 (link) Sera from 3 mice immunized with MPER and Co-PoP liposomes was pooled and IgG was isolated using immobilized Protein G beads (VWR # PI20398) according to vendor protocol. 2F5 was graciously provided by the free NIH AIDS reagent program. 1×104 TZM-bl receptor cells per well were plated in a 96-well plate the day before infection. HIV (multiplicity of infection of 0.1) was incubated with antibodies for 30 min at 37 °C, added to the cells and spinoculated at 1000×g for 1 h at 25 °C followed by further incubation for 2 days at 37 °C. Cell viability and viral entry were respectively assessed using the CellTiter-Fluor (Promega) and One-Glo (Promega) assays according to manufacturer protocol.
Protein g beads
Manufactured by Avantor
Protein G beads are a type of affinity chromatography material used for the purification of antibodies. They consist of Protein G, a bacterial cell wall protein, covalently coupled to agarose beads. Protein G has a high affinity for the Fc region of immunoglobulins, allowing it to effectively capture and purify antibodies from complex mixtures.
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2 protocols using protein g beads
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Viral Entry Neutralization Assay
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Viral Entry Neutralization Assay
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Viral entry experiments were carried out as previously described.49 (link) Sera from 3 mice immunized with MPER and Co-PoP liposomes was pooled and IgG was isolated using immobilized Protein G beads (VWR # PI20398) according to vendor protocol. 2F5 was graciously provided by the free NIH AIDS reagent program. 1×104 TZM-bl receptor cells per well were plated in a 96-well plate the day before infection. HIV (multiplicity of infection of 0.1) was incubated with antibodies for 30 min at 37 °C, added to the cells and spinoculated at 1000×g for 1 h at 25 °C followed by further incubation for 2 days at 37 °C. Cell viability and viral entry were respectively assessed using the CellTiter-Fluor (Promega) and One-Glo (Promega) assays according to manufacturer protocol.
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