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Non-affinity purification of a nanobody by void-exclusion anion exchange chromatography and multimodal weak cation exchange chromatography

separation and purification Technology. 2019; 
XiyingFanaYuhangZhoubJianliYuaFeiLibQuanChenaHaipengSongbDongxiaoFengcWenshuaiLiuaRuiNianaMoXiana
Products/Services Used Details Operation
Molecular Biology Reagents The immunoenzymetric assay for the measurement of E. coli HCP was purchased from Cygnus Technologies Inc. (cat. F410, Southport, USA). The ToxinSensor™ Chromogenic Limulus Amebocyte Lysate Endotoxin Assay Kit was obtained from GenScript (cat. L00350C, Nanjing, China). 12% Mini-Protein Tgxtm precast gels were purchased from GenScript (cat. M01212C and M01215C, Nanjing, China), MOPS electrophoresis buffer powder was obtained from Genscript (cat. M00138). The PageRuler Prestained Protein Ladder was purchased from Thermo Fisher Scientific (cat. 26616, Rockford, USA).  Get A Quote

摘要

Multimodal chromatography resins have been reported to be excellent alternatives to classical absorbents in the purification of antibodies. In this study, void-exclusion anion exchange chromatography (VEAX) and electropositive multimodal chromatography (Capto MMC) were integrated for tag-free nanobody (2D5) purification. VEAX was systematically characterized for its potential use in the initial 2D5 separation. VEAX achieved a 89.9% reduction in host cell proteins (HCPs), a 99.6% reduction in DNA, a 99.9% reduction in endotoxins, and a 100% reduction in aggregates from periplasmic proteins, with a 2D5 recovery of near 100% and a purity enhancement from 3.5% to 25.0%. Subsequently, Capto MMC chromatography was se... More

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