Laboratory of Molecular Biology and Biotechnology, Department of Biochemistry, San Luis University, Ejército de los Andes 950, D5700HHW, San Luis, Argentina.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Sep 15;879(26):2741-5. doi: 10.1016/j.jchromb.2011.07.040. Epub 2011 Aug 6.
The objective of this study was the development of affinity adsorbent particles with the appropriate characteristics to be applied in protein purification using the affinity ultrafiltration method. To prepare affinity macroligands Cibacron Blue 3GA, as a ligand molecule, was immobilized by covalent bonding onto yeast cell walls, the support material or matrix. The maximum attachment of the ligand to the matrix was 212 μmol/g (ligand dry weight/yeast dry weight). Lysozyme was selected as the protein model for the adsorption studies. Its adsorption onto the matrix without ligand and matrix with attached ligand were investigated batch-wise. The adsorption equilibrium isotherms appeared to follow a typical Langmuir isotherm. The maximum adsorption capacity (q(m)) of the Cell-Cibacron macroligand for lysozyme was 110 mg/ml of wet macroligand. The adsorbent was also employed for the separation of lysozyme from hen egg white. High purity lysozyme was obtained.
本研究的目的是开发具有适当特性的亲和吸附剂颗粒，以便在使用亲和超滤法进行蛋白质纯化时应用。为了制备亲和大环配体 Cibacron Blue 3GA，作为配体分子，通过共价键固定在酵母细胞壁、载体材料或基质上。配体与基质的最大附着量为 212 μmol/g（配体干重/酵母干重）。溶菌酶被选为吸附研究的蛋白质模型。研究了未结合配体的基质和结合了配体的基质对其的分批吸附。吸附平衡等温线似乎遵循典型的 Langmuir 等温线。Cell-Cibacron 大环配体对溶菌酶的最大吸附容量（q(m)）为 110 mg/ml 湿大环配体。该吸附剂还用于从鸡蛋蛋清中分离溶菌酶。获得了高纯度的溶菌酶。
