IMMUNOAFFINITY CHROMATOGRAPHY OF LYMPHOCYTE SUBPOPULATIONS USING TERT-AMINE DERIVED MATRICES WITH ADSORBED ANTIBODIES
IMMUNOAFFINITY CHROMATOGRAPHY OF LYMPHOCYTE SUBPOPULATIONS USING TERT-AMINE DERIVED MATRICES WITH ADSORBED ANTIBODIES
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DOI：
10.1016/0142-9612(88)90087-7
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发表时间：
1988-05-01
期刊：
影响因子：
14
通讯作者：
TSURUTA, T
中科院分区：
文献类型：
--
作者：
KATAOKA, K;SAKURAI, Y;TSURUTA, T
制备了用于细胞亲和层析的新型聚合物固相基质，并强调了其与传统基质相比的优点。这些新基质是聚甲基丙烯酸2-羟乙基酯（PHEMA）的衍生物，含有少量的氨基化合物作为共聚单体。将其应用于大鼠肠系膜淋巴结IgG+和IgG-淋巴细胞的免疫亲和选择。简单的物理吸附足以将抗大鼠IgG抗体固定在这些基质上，从而使我们省去了抗体在基质上共价连接的繁琐程序。由于基质本身对淋巴细胞的非特异性吸附极低，因此非常稀释的抗体溶液（0.02-0.08 mg/ml）就足以进行柱调理。该分离方法可在7分钟内获得纯度超过90%的IgG-淋巴细胞，收率接近95%。此外，通过轻移基质，从柱上回收吸附细胞部分，获得了产量较高的IgG+淋巴细胞（负载数的80-90%）。
New polymeric solid-phase matrices for cell affinity chromatography were prepared and their advantageous characteristics compared with conventional matrices were highlighted. These new matrices are derivatives of poly(2-hydroxyethyl methacrylate) (PHEMA) containing a slight quantity of amino compounds as a co-monomer. They were applied to immunoaffinity selection between IgG+ and IgG- lymphocytes of the rat mesenteric lymph node. Simple physical adsorption was sufficient for anti-rat IgG antibodies to be immobilized on these matrices, allowing us to omit the laborious procedure of covalent-linking of antibodies on a matrix. As the matrices themselves showed extremely low non-specific adsorption of lymphocytes, a very dilute solution of antibody (0.02-0.08 mg/ml) was enough for column conditioning. This separation method gave IgG- lymphocytes of more than 90% purity and almost 95% yield within as short a time as 7 min. Further, IgG+ lymphocytes were obtained in good yield (80-90% of loaded number) by recovering the adsorbing cell fraction from the column by gentle pipetting of the matrix.