Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris

被引:36
作者
Ohtani, W [1 ]
Nawa, Y [1 ]
Takeshima, K [1 ]
Kamuro, H [1 ]
Kobayashi, K [1 ]
Ohmura, T [1 ]
机构
[1] Green Cross Co, Div Res, Osaka 573, Japan
关键词
recombinant human serum albumin; fatty acids; viscosity; colloid osmotic pressure; antigenicity;
D O I
10.1006/abio.1997.2480
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 how, 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). (C) 1998 Academic Press.
引用
收藏
页码:56 / 62
页数:7
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