Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris.
Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris.
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DOI：
10.1006/abio.1997.2480
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发表时间：
1998-02
影响因子：
2.9
通讯作者：
W. Ohtani;Y. Nawa;K. Takeshima;H. Kamuro;K. Kobayashi;T. Ohmura
中科院分区：
文献类型：
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作者：
W. Ohtani;Y. Nawa;K. Takeshima;H. Kamuro;K. Kobayashi;T. Ohmura
分析比较了巴斯德毕赤酵母重组人血清白蛋白(RHSA)和血浆来源的人血清白蛋白(PHSA)的理化和免疫化学性质。在pH值为6.7+/-0.1的胶体渗透压下测得rHSA的第二维里系数与PHSA的第二维里系数无显著差异(P>；0.05)。25%重组人血清白蛋白溶液表现出牛顿流动，在20℃+/-0.02℃时，25%人血清白蛋白溶液的粘度与25%PHSA溶液的粘度没有显著差异(P>；0.05)。以9-蒽基重氮甲烷为荧光标记剂，采用反相高效液相色谱法分析重组人血清白蛋白的长链和中链脂肪酸组成。PHSA的脂肪酸总量高于RHSA。重组人血清白蛋白制剂的脂肪酸组成与PHSA制剂相同。而重组人血清白蛋白中棕榈酸(C16：0)和硬脂酸(C18：0)的含量明显低于PHSA。有趣的是，我们发现巴斯德毕赤酵母产生亚麻酸(C18：3)，因为在重组人血清白蛋白中检测到亚麻酸。用抗PHSA多克隆抗体的平行线分析法检测rHSA的免疫化学性质，结果与PHSA一致(P>；0.05)。
We analyzed and compared the physicochemical and immunochemical properties of recombinant human serum albumin (rHSA) from Pichia pastoris with those of plasma-derived human serum albumin (pHSA). The second virial coefficient of rHSA, obtained from colloid osmotic pressure measurements at pH 6.7 +/- 0.1 was not significantly different from that of pHSA (P > 0.05). A 25% rHSA solution exhibited Newtonian flow, and the viscosity of 25% rHSA at 20 +/- 0.02 degrees C was not significantly different from that of 25% pHSA (P > 0.05). We analyzed the long- and medium-chain fatty acid composition of rHSA by reverse-phase HPLC using 9-anthryldiazomethane as the fluorescent labeling reagent. The total amount of fatty acid was higher for pHSA than for rHSA. The fatty acid composition of the rHSA preparation was the same as that of the pHSA preparation. However, the amounts of palmitic acid (C16:0) and stearic acid (C18:0) in rHSA were much lower than those in pHSA. Interestingly, we found that P. pastoris produced linolenic acid (C18:3) because it was detected in rHSA. The immunochemical properties of rHSA were analyzed by a parallel line assay method using anti-pHSA polyclonal antibody, and were identical to those of pHSA (P > 0.05).