Boughton B J, Simpson A, Wharton C
Vox Sang. 1984;46(5):254-9. doi: 10.1111/j.1423-0410.1984.tb00084.x.
Using human peripheral blood monocytes to assay opsonic protein activity, we have examined the efficiency of various procedures for isolating fibronectin from plasma for experimental or therapeutic use. In addition, we have assessed the protein's opsonic activity after cold storage, and after heat treatment to inactivate hepatitis virus. The purification procedures recovered only 30% of available plasma fibronectin whilst cold storage and heat treatment of the purified protein removed all of its remaining opsonic activity. This was associated with no alteration in overall molecular weight or in subunit size but was accompanied by changes in ultraviolet spectrum, suggesting a conformational change in the protein structure. Initial experiments to protect the purified protein against these changes were unsuccessful and unless further attempts are more encouraging, fresh-frozen plasma may be the only current economic source of opsonically active fibronectin. Since this would waste other valuable proteins required for other purposes, the widespread use of plasma fibronectin outside of clinical trials may be unjustified at this present time.
我们利用人外周血单核细胞来检测调理素蛋白活性，研究了从血浆中分离纤连蛋白用于实验或治疗的各种方法的效率。此外，我们评估了纤连蛋白在冷藏后以及热处理灭活肝炎病毒后的调理活性。纯化过程仅回收了血浆中30%的纤连蛋白，而对纯化后的蛋白进行冷藏和热处理后，其剩余的所有调理活性均丧失。这与整体分子量或亚基大小无变化相关，但伴随着紫外光谱的改变，提示蛋白结构发生了构象变化。保护纯化蛋白免受这些变化影响的初步实验未成功，除非进一步的尝试更令人鼓舞，否则新鲜冷冻血浆可能是目前唯一具有调理活性的纤连蛋白的经济来源。由于这会浪费其他用途所需的其他有价值的蛋白质，目前在临床试验之外广泛使用血浆纤连蛋白可能不合理。
