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单克隆抗体的产生和筛选;作者给12周龄Lewis雌性大鼠经ip和sc注射1.0×10~7培养前-B细胞系70Z/3,进行初次和加强免疫。间隔4个月,再用BALB/c小鼠出生第1天肝细胞7×10~7进行初次和加强免疫,末次注射后4天,取免疫鼠脾淋巴细胞与sP2/0-Mg14细胞,采用Mc Kearn等人的方法进行细胞融合。阳性孔细胞在半固体琼脂中进行细胞克隆化。细胞结合放射免疫测定(CRIA)按Longenecker等人方案,细胞单色与双色免疫荧光技术按Loken等人方法进行;用荧光-激活细胞分类器(FACS Ⅱ)来测定各单个细胞的荧光和光散射信号(0.7°—15°)。
Monoclonal antibody production and screening; the author Lewis rats 12 weeks of age ip and sc injected 1.0 × 10-7 pre-cultured B cell line 70Z / 3, the first and booster immunization. Four months after the first immunization, BALB / c mice were immunized for 7 × 10 ~ 7 days on day 1 with primary and booster immunization. Spleen lymphocytes and sP2 / 0-Mg14 cells were immunized 4 days after the last injection. Mc Kearn et al.’s method for cell fusion. Positive well cells were cloned in semi-solid agar. Cell-bound radioimmunoassay (CRIA) The Longenecker et al. Protocol, cell monocolor and two-color immunofluorescence techniques were performed by the method of Loken et al. Fluorescence-activated cell sorter (FACS II) was used to determine the fluorescence and light scattering signals of individual cells (0.7 ° -15 °).