METHODS FOR SEPARATION PURIFICATION AND MEASUREMENT OF 9 COMPONENTS OF HEMOLYTIC COMPLEMENT IN GUINEA-PIG SERUM
METHODS FOR SEPARATION PURIFICATION AND MEASUREMENT OF 9 COMPONENTS OF HEMOLYTIC COMPLEMENT IN GUINEA-PIG SERUM
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DOI：
10.1016/0019-2791(66)90292-8
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发表时间：
1966-01-01
期刊：
影响因子：
--
通讯作者：
TAMURA, N
中科院分区：
文献类型：
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作者：
NELSON, RA;JENSEN, J;TAMURA, N
C[image]1在离子强度0.04和pH 7.5时从正常豚鼠血清中沉淀。可在透析过程中用0[中心点]005 M磷酸盐缓冲液（pH 7.5）再沉淀进一步纯化。C[image]3b在离子强度0.04-0.06和pH 5.0时沉淀。在离子强度约为0.12时，它很容易通过从DEAE-纤维素中洗脱而纯化。C[image]3c在离子强度0.02和pH 5.5时沉淀。它可以通过在约0.065离子强度下从DEAE-纤维素洗脱来纯化。当（NH 4）2SO 4加入到约2.2 M时，C[image]2和C[image]3d保持可溶，然后可以在约3.0 M（NH 4）2SO 4下沉淀。这两种组分可通过CM-纤维素色谱法容易地分离。8种组分从CM-纤维素中析出的顺序和近似离子强度为C[图像]2，0.05; C[图像]3f，0.12; C[图像]4，0.13; C[图像]3c，0.135; C[图像]3b，0.14; C[图像] 3e，0.145; C[图像]3a，0.18;和C[图像]3d，0.20。从DEAE-纤维素洗脱的顺序和离子强度为：C[图像]3e，0.03; C[图像]3a，0.035; C[图像]3c和C[图像]3f，0.065; C[图像]3d，0.095; C[图像]3b和C[图像]4，0.12。从DEAE-纤维素中得到C[image]2的产率一直很低。在正常豚鼠血清中，使中间复合物EAb C[image] 1，4，2，3c、EAb C [image] 1，4，3c或EAb C[image]4，3c上的C [image]3c位点失活的物质在约0.05至0.06离子强度下从DEAE-纤维素洗脱。另一种使C[image]1失活的物质在离子强度约为0.12时从DEAE-纤维素中洗脱出来。
C[image]1 precipitates from normal guinea pig serum at ionic strength 0.04 and pH 7.5. It may be purified further by reprecipitation during dialysis against 0[center dot]005 M phosphate buffer, pH 7.5. C[image]3b precipitates at ionic strength 0.04-0.06 and pH 5.0. It is easily purified by elution from DEAE-cellulose at about 0.12 ionic strength. C[image]3c precipitates at ionic strength 0.02 and pH 5.5. It can be purified by elution from DEAE-cellulose at about 0.065 ionic strength. C[image]2 and C[image]3d remain soluble when (NH4)2SO4 is added to about 2.2 M and then can be precipitated at about 3.0 M (NH4)2SO4. These two components can be separated easily by chromatography on CM-cellulose. The sequence and the approximate ionic strength at which 8 components elute from CM-cellulose is C[image]2, 0.05; C[image]3f, 0.12; C[image]4, 0.13; C[image]3c, 0.135; C[image]3b, 0.14; C[image]3e, 0.145; C[image]3a, 0.18; and C[image]3d, 0.20. The sequence and ionic strength of elution from DEAE-cellulose is: C[image]3e, 0.03; C[image]3a, 0.035; C[image]3c and C[image]3f, 0.065; C[image]3d, 0.095; C[image]3b and C[image]4, 0.12. The yield of C[image]2 from DEAE-cellulose has been low. A substance in normal guinea pig serum which inactivates the C [image]3c site on the intermediate complexes, EAb C[image]l,4,2,3c, EAb C[image]l,4,3c or EAb C[image]4,3c, elutes from DEAE-cellulose at about 0.05 to 0.06 ionic strength. Another substance which inactivates C[image]1 elutes from DEAE-cellulose at about 0.12 ionic strength.