ISOLATION OF PURE IGG1, IGG2A AND IGG2B IMMUNOGLOBULINS FROM MOUSE SERUM USING PROTEIN A-SEPHAROSE
ISOLATION OF PURE IGG1, IGG2A AND IGG2B IMMUNOGLOBULINS FROM MOUSE SERUM USING PROTEIN A-SEPHAROSE
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DOI：
10.1016/0161-5890(78)90070-6
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发表时间：
1978-01-01
期刊：
影响因子：
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通讯作者：
JENKIN, CR
中科院分区：
文献类型：
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作者：
EY, PL;PROWSE, SJ;JENKIN, CR
本文报道了一种简便、快速、产率接近100%的小鼠IgG_1、IgG_2a和IgG_2b纯品的分离方法。将小鼠血清在蛋白质A-Sepharose 4 B上分级分离，并测量免疫球蛋白的回收率作为pH值的函数。当将血清施加到pH 8.0的柱上时，IgM，伊加和IgE几乎定量地回收在流出物中与非免疫球蛋白血清组分一起。如果不超过柱的结合能力，则基本上所有IgG都保留在pH 8.0下，并且这不能通过洗涤洗脱。用pH逐渐降低的缓冲液，分别在pH 6.0-7.0、pH 4.5-5.0和pH 3.5-4.0下依次洗脱IgG 1、IgG 2a和IgG 2b。
A simple and rapid method for isolating pure mouse IgG1, IgG2aand IgG2bimmunoglobulins in nearly 100% yield is described. Mouse serum was fractionated on protein A-Sepharose 4B and the recovery of immunoglobulins was measured as a function of pH. When serum was applied to the column at pH 8.0, IgM, IgA and IgE were almost quantitatively recovered in the effluent together with non-immunoglobulin serum components. Providing the binding capacity of the column was not exceeded, essentially all IgG was retained at pH 8.0 and this could not be eluted by washing. Using buffers of decreasing pH, IgG1, IgG2aand IgG2bwere sequentially eluted at pH 6.0–7.0, pH 4.5–5.0 and pH 3.5–4.0, respectively.