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  • Strategies for the one-step...
    Barbosa, Oveimar; Ortiz, Claudia; Berenguer-Murcia, Ángel; Torres, Rodrigo; Rodrigues, Rafael C.; Fernandez-Lafuente, Roberto

    Biotechnology advances, 09/2015, Letnik: 33, Številka: 5
    Journal Article

    In this review, we detail the efforts performed to couple the purification and the immobilization of industrial enzymes in a single step. The use of antibodies, the development of specific domains with affinity for some specific supports will be revised. Moreover, we will discuss the use of domains that increase the affinity for standard matrices (ionic exchangers, silicates). We will show how the control of the immobilization conditions may convert some unspecific supports in largely specific ones. The development of tailor-made heterofunctional supports as a tool to immobilize–stabilize–purify some proteins will be discussed in deep, using low concentration of adsorbent groups and a dense layer of groups able to give an intense multipoint covalent attachment. The final coupling of mutagenesis and tailor made supports will be the last part of the review. •Use of immobilized antibodies to get the one step immobilization–purification•Use of domains with specific affinity for some ligands to immobilize/purify tagged proteins•Immobilization/purification by using of domains that increase the enzyme affinity for standard supports•Glyoxyl supports and one step immobilization–purification of multimeric proteins•Medium engineering and standard supports for immobilization/purification•Tailor made heterofunctional supports for large protein immobilization/purification