ZCa: A Protein A-Derived Domain with Calcium-Dependent Affinity for Mild Antibody Purification
| Autor: | Sophia Hober, Sara Kanje, Julia Scheffel |
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| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Modern medicine Chromatography biology Elution 010401 analytical chemistry chemistry.chemical_element Calcium Ligand (biochemistry) 01 natural sciences 0104 chemical sciences 03 medical and health sciences chemistry Tetramer Affinity chromatography biology.protein Antibody Protein A 030304 developmental biology |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071607749 |
| DOI: | 10.1007/978-1-0716-0775-6_17 |
| Popis: | Therapeutic antibodies are at the forefront of modern medicine where high purity, which is typically obtained by Protein A-based affinity purification, is of utmost importance. In this chapter, we present a method for neutral and selective purification of antibodies by utilizing an engineered affinity ligand, ZCa, derived from Protein A. This domain displays a calcium-dependent binding of antibodies and has been multimerized and immobilized to a chromatography resin to achieve an affinity matrix with high binding capacity. IgG antibodies can be eluted from the tetrameric ZCa ligand at pH 7 with the addition of EDTA, or at pH 5.5 with EDTA for purification of monoclonal IgG1, which is significantly milder than the low pH (3-4) required in conventional Protein A affinity chromatography. Here, a protocol for selective capture of IgG with elution at neutral pH from a ZCa tetramer ligand immobilized on a chromatography resin is described. |
| Databáze: | OpenAIRE |
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