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Recombinant β-Glucocerebrosidase specific immunoaffinity ligands selected from phage-displayed combinatorial scFv libraries.
Protein Expression and Purification ( IF 1.4 ) Pub Date : 2020-01-22 , DOI: 10.1016/j.pep.2020.105573 R L Anisimov 1 , O A Ershova 1 , A V Ershov 1 , M A Filatova 1 , S A Katorkin 1 , V M Simonov 1
Protein Expression and Purification ( IF 1.4 ) Pub Date : 2020-01-22 , DOI: 10.1016/j.pep.2020.105573 R L Anisimov 1 , O A Ershova 1 , A V Ershov 1 , M A Filatova 1 , S A Katorkin 1 , V M Simonov 1
Affiliation
Antibodies specific to β-Glucocerebrosidase were selected from phage displayed naïve scFv libraries. Biopannings were performed against recombinant human protein β-Glucocerebrosidase immobilized on polystyrene surface, specific phages were eluted with 50% ethylene glycol in citrate buffer (pH 6.0). Several specific binders were discovered and converted to full-size hIgG1 antibodies leading to highly stable binders with dissociation constants (Kd) in the range 10-40 nM. The antibodies were used further as ligands for affinity chromatography, where efficient and selective recovery of biologically active β-Glucocerebrosidase from cultured media of Chinese hamster ovary cells was demonstrated. β-Glucocerebrosidase was purified to nearly homogeneous state and had specific activity comparable to the commercially available preparations (40-44 U/mg protein). The obtained immunoaffinity sorbents have high capacity and can be easily regenerated.
更新日期：2020-01-22
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