Non-specific binding of proteins by substituted agaroses.
Non-specific binding of proteins by substituted agaroses.
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取代琼脂糖对蛋白质的非特异性结合。
DOI：
10.1007/978-1-4684-6982-0_4
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发表时间：
1974
影响因子：
--
通讯作者：
B. Hofstee
中科院分区：
文献类型：
--
作者：
B. Hofstee
吸附剂的取代度由卵清蛋白或丽春红S的”不可逆”结合能力确定，如结果中所述。该节还描述了用于此目的的滴定法。通过分别向洗脱液中加入盐或乙二醇的影响，确定吸附剂与蛋白质结合的极性（如静电）或非极性（疏水）方面的相对程度（1-3）。对于几个实验，特别是那些通过盐梯度进行洗脱的实验，使用自动化程序，其中用蠕动泵将洗脱液泵入柱中，并通过测量280或225 nm处的UV吸光度连续监测洗脱液的蛋白质含量。然而，手动测量吸光度的逐步洗脱具有优于自动化程序的优点，因为可以同时操作大量的柱。程序的细节在附图和表格的图例中描述。
MethodsThe degrees of substitution of the adsorbents were determined from the capacities for" irreversible" binding of ovalbumin or of Ponceau S, as described under RESULTS. A titrimetric method for this purpose also is described in that section.The relative degree of the polar (eg, electrostatic) or apolar (hydrophobic) aspects of the binding of a protein by an adsorbent was determined by the effects of the addition to the eluant of salt or of ethylene glycol, respectively (1-3). For several experiments, particularly those in which elution was carried out by means of a salt gradient, an automated procedure was used whereby the eluant was pumped into the column with a peristaltic pump and the protein content of the eluate was monitored continuously through measurement of the UV light absorbance at either 280 or 225 nm. However, stepwise elution with manual measurement of absorbance of the eluate has the advantage over automated procedures in that a large number of columns can be operated simultaneously. Details of the procedures are described in the legends to the Figures and the Table.