The Key Laboratory of Leather Chemistry and Engineering of Ministry of Education, Sichuan University, Chengdu 610065, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Jun 1;928:131-8. doi: 10.1016/j.jchromb.2013.03.031. Epub 2013 Apr 8.
The separation of proteins is a key step in biomedical and pharmaceutical industries. In the present investigation, the collagen fiber adsorbent (CFA) was exploited as column packing material to separate proteins. Bovine serum albumin (BSA), bovine hemoglobin (Hb) and lysozyme (LYS) that have different isoelectric points (pIs) were selected as model proteins to investigate the separation ability of CFA to proteins. In batch adsorption, the adsorption behaviors of these proteins on CFA under different pHs and ionic strengths indicated that the electrostatic interaction plays a predominant role in the adsorption of proteins on CFA. CFA exhibited high adsorption capacity to Hb and LYS. In column separation, the proteins were completely separated by adjusting pH and ionic strength of the eluent. The increase of flow rate could reduce the separation time with no influence on the recovery of protein in the experimental range. The protein recovery was higher than 90% even when the CFA column was re-used for 4 times in separation of BSA and LYS, and the retention time of BSA or LYS was almost constant during the repeated applications. In addition, as a practical application, LYS was successfully separated from chicken egg white powder by CFA column.
蛋白质分离是生物医药和制药行业的关键步骤。在本研究中，我们利用胶原纤维吸附剂（CFA）作为柱填料来分离蛋白质。选择具有不同等电点（pI）的牛血清白蛋白（BSA）、牛血红蛋白（Hb）和溶菌酶（LYS）作为模型蛋白，以研究 CFA 对蛋白质的分离能力。在批量吸附实验中，在不同 pH 值和离子强度下，这些蛋白质在 CFA 上的吸附行为表明，静电相互作用在蛋白质在 CFA 上的吸附中起主要作用。CFA 对 Hb 和 LYS 具有较高的吸附能力。在柱分离中，通过调节洗脱液的 pH 值和离子强度可以完全分离蛋白质。在实验范围内，增加流速可以缩短分离时间，而不会影响蛋白质的回收率。即使在 BSA 和 LYS 的分离中重复使用 CFA 柱 4 次，蛋白质的回收率也高于 90%，并且在重复使用过程中 BSA 或 LYS 的保留时间几乎保持不变。此外，作为实际应用，通过 CFA 柱成功地从鸡蛋白粉中分离出 LYS。
