Effective elution of antibodies by arginine and arginine derivatives in affinity column chromatography
Effective elution of antibodies by arginine and arginine derivatives in affinity column chromatography
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DOI：
10.1016/j.ab.2005.07.004
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发表时间：
2005-10-15
影响因子：
2.9
通讯作者：
Arakawa, T
中科院分区：
文献类型：
--
作者：
Ejima, D;Yumioka, R;Arakawa, T
结果表明，以精氨酸为洗脱剂，可显著提高从蛋白A亲和层析中回收单体抗体的效率。为了扩大精氨酸在抗体纯化中的应用，并深入了解精氨酸洗脱的机理，我们在蛋白A层析中比较了精氨酸与柠檬酸盐、盐酸胍(GdnHCl)、精氨酸衍生物以及其他氨基酸的差异。我们还将精氨酸应用于抗原亲和层析中多克隆抗体的洗脱。如前所述，精氨酸可以有效地洗脱单抗IgG1和IgG4。两种精氨酸衍生物，乙酰精氨酸和胍丁胺，在pH 4.0或更高时能有效洗脱，这与精氨酸相当。另一方面，其他氨基酸，如甘氨酸、脯氨酸、赖氨酸和组氨酸，在相同的pH条件下比精氨酸的效果要差得多。精氨酸浓度越高，洗脱效果越好，但在pH为4.3时，柠檬酸的洗脱效果不明显。精氨酸在抗原偶联亲和层析柱分离PABS中也是有效的。虽然GdnHCl在类似的条件下也是有效的，但洗脱的物质比精氨酸洗脱的蛋白质表现出更多的聚集。(C)2005 Elsevier Inc.保留所有权利。
It has been shown that the recovery of monomeric antibodies from protein A affinity chromatography is enhanced significantly by using arginine as an eluent. To extend the applications of arginine to antibody purification and obtain an insight into the mechanism of arginine elution, we compared arginine with citrate, guanidine hydrochloride (GdnHCl), arginine derivatives, and other amino acids in protein A chromatography. We also applied arginine to elution of polyclonal antibodies (pAbs) in antigen affinity chromatography. As described previously, arginine was effective in eluting monoclonal antibodies IgG1 and IgG4. Two arginine derivatives, acetyl-arginine and agmatine, resulted in efficient elution at pH 4.0 or higher, and this was comparable to arginine. On the other hand, other amino acids, such as glycine, proline, lysine, and histidine, are much less effective than arginine under identical pH conditions. Whereas elution increased with arginine concentration, elution with citrate was insignificant in excess of I M at pH 4.3. Arginine was also effective in fractionation of pAbs using antigen-conjugated affinity columns. Although GdnHCl was also effective under similar conditions, the eluted material showed more aggregation than did the protein eluted by arginine. (C) 2005 Elsevier Inc. All rights reserved.