Author: Bancroft, Tara; DeBuysscher, Blair L.; Weidle, Connor; Schwartz, Allison; Wall, Abigail; Gray, Matthew D.; Feng, Junli; Steach, Holly R.; Fitzpatrick, Kristin S.; Gewe, Mesfin M.; Skog, Patrick D.; Doyle-Cooper, Colleen; Ota, Takayuki; Strong, Roland K.; Nemazee, David; Pancera, Marie; Stamatatos, Leonidas; McGuire, Andrew T.; Taylor, Justin J.
Title: Detection and activation of HIV broadly neutralizing antibody precursor B cells using anti-idiotypes Document date: 2019_10_7
ID: 63yvpuqx_61
Snippet: Fab and cleavable single-chain variable fragment (c/scFv) expression, purification, and complex preparation IB2 and IB3 IgG purified from hybridomas were digested over activated Ficin on Agarose Resin (Thermo Fisher Scientific) for 48 h to obtain Fabs. Fabs for iglb12 were generated by digesting with 1 µg Lys C (Roche) for 10 mg of IgG overnight at 37°C. The Fabs were purified from Fc regions by collecting the fraction that did not bind to a Pr.....
Document: Fab and cleavable single-chain variable fragment (c/scFv) expression, purification, and complex preparation IB2 and IB3 IgG purified from hybridomas were digested over activated Ficin on Agarose Resin (Thermo Fisher Scientific) for 48 h to obtain Fabs. Fabs for iglb12 were generated by digesting with 1 µg Lys C (Roche) for 10 mg of IgG overnight at 37°C. The Fabs were purified from Fc regions by collecting the fraction that did not bind to a Protein A column (Thermo Fisher Scientific) and by size exclusion chromatography using Superdex 200 (GE Healthcare Life Sciences) in 5 mM Hepes and 150 mM NaCl.
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