title: Preparation and characterization of novel IgG affinity resin coupling anti-Fc camelid single-domain antibodies.
abstract: This work aimed to evaluate novel affinity resin used to purify immunoglobulin G (IgG) with a variable domain of the heavy chain of the heavy-chain antibody (VHH) as an affinity ligand. The VHH, isolated from a naïve camelid single-domain phage display library, exhibits not only affinity to the fragment crystallizable (Fc) region of IgG but also high thermal stability. This anti-Fc VHH (AFV) was expressed as a soluble protein in Escherichia coli and purified using a simple heat treatment procedure. The effects of pH and NaCl concentrations on the capacity of AFV resin were also investigated. Results showed a robust property of the AFV resin. It could bind IgGs at various pH conditions (from 6.0 to 9.0) and NaCl concentrations. The static binding capacities of AFV resin ranged from 3.40±0.53mg/ml to 15.04±0.37mg/ml measured using rabbit, mouse, and human IgGs. The bound IgGs can be efficiently eluted at pH 5.0, which is conducive to acid-sensitive IgGs and prevents the aggregation of IgGs. After 10 purification cycles or a 7-day period of storage at 37°C, recovery did not decrease. These findings suggested that VHHs from non-immunized library could also be robust and functional reagent as an affinity purification ligand.
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China.
authors.forename: Zhui
authors.initials: Z
authors.lastname: Tu
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, China; Shanxi Defcred Biotechnology Co., Ltd., Shanxi, China. Electronic address: fujinheng@ncu.edu.cn.
authors.forename: Jinheng
authors.initials: J
authors.lastname: Fu
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, China.
authors.forename: Zhibing
authors.initials: Z
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Shanxi Defcred Biotechnology Co., Ltd., Shanxi, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.affiliation: Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Tao
mesh_terms: D000818:Animals; D002021:Buffers; D002161:Camelids, New World; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D055550:Protein Stability; D011961:Receptors, Fc / Q000378:metabolism*; D011994:Recombinant Proteins / Q000302:isolation & purification; D012117:Resins, Synthetic / Q000737:chemistry*; D061905:Single-Domain Antibodies / Q000276:immunology*; D012995:Solubility; D013696:Temperature
chemical_list: D002021:Buffers; D007074:Immunoglobulin G; D011961:Receptors, Fc; D011994:Recombinant Proteins; D012117:Resins, Synthetic; D061905:Single-Domain Antibodies
keywords: Affinity chromatography; Anti-Fc; Purification of IgG; Single domain antibody