Departments of Bio-Resource Engineering, Vancouver, British Columbia, V6T 1Z4, Canada.
Biotechnol Bioeng. 1992 Dec 20;40(11):1388-94. doi: 10.1002/bit.260401113.
An automated liquid chromatography system was developed to carry out the separation of an egg yolk immunoglobulin (IgY) using cation exchange media. Industrially separated egg yolk was diluted 10 times with distilled water, the pH adjusted to 5.5, and the water-soluble protein fraction separated from lipoproteins by sedimentation. The supernatant was filtered and then applied to a column packed with a cation exchanger within an automated liquid chromatography system. Different operating conditions were investigated using phosphate buffer in order to assess the effect on recovery and purity. Fractions as pure as 80% could be collected and a recovery of the chromatography step of about 65% was obtained for a purity of 60% using either a linear or step gradient. The overall recovery for the process was 34% if one-step dilution/extraction is used for lipoprotein separation by sedimentation, and 51% if two-step dillution/extraction is used. Further improvement of the yield to about 60% is possible using centrifugation for lipoprotein separation. The automated system confers many advantages, the key elements being the time savings and accurate control of the process.
建立了一个自动化液相色谱系统，采用阳离子交换介质分离蛋黄免疫球蛋白（IgY）。工业分离的蛋黄用蒸馏水稀释 10 倍，将 pH 值调至 5.5，并通过沉降将水溶性蛋白质部分与脂蛋白分离。将上清液过滤，然后应用于填充在自动化液相色谱系统内的阳离子交换柱。使用磷酸盐缓冲液研究不同的操作条件，以评估其对回收率和纯度的影响。可以收集纯度高达 80%的馏分，并且使用线性或分步梯度，对于纯度为 60%，色谱步骤的回收率约为 65%。如果使用沉降进行脂蛋白的一步稀释/提取，则整个过程的总回收率为 34%；如果使用两步稀释/提取，则回收率为 51%。使用离心法分离脂蛋白可进一步提高产率至约 60%。自动化系统具有许多优点，关键要素是节省时间和准确控制过程。
