LAK细胞中一种自由基清除剂——SLP的分离纯化与抗体制备

Purification of a Radical Scavenger——SLP from LAK Cells and Preparation of Antibody Against SLP *

ＳＯＤ样蛋白（ＳＯＤ－ｌｉｋｅ ｐｒｏｔｅｉｎ，ＳＬＰ）是从ＬＡＫ 细胞中发现的一种具有自由基清除功能的蛋白．为阐明ＳＬＰ的生化特性和确定其蛋白序列或基因序列，采用ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ ＦＦ层析从ＬＡＫ 细胞中得到部分纯化的ＳＬＰ，并采用ＩＥＦ等电聚焦电泳，活性染色，将含有活性蛋白的胶条直接免疫Ｂａｌｂ／ｃ 小鼠，制备抗血清并筛选得到多株有中和ＳＬＰ清除自由基活性的单克隆抗体．Ｗｅｓｔｅｒｎ ｂｌｏｔ结果表明ＳＬＰ分子量约为６７ ｋＤ，并测得ｐＩ约为４．２．

SLP,a SOD like protein that can scavenge O -· 2 free radical effectively is found in LAK cells. To elucidate its biochemical properties and identify the protein sequence or gene sequence,SLP was partially purified from human LAK cells with DEAE Sepharose FF chromatography.The partial purified SLP was electrophoresised with IEF to obtaining its antibodies,and negative staining method was used for SLP activity determination.The gels of negative staining band were cut and embed into Balb/c mice subcutaneously for three times.Monoclonal and polyclonal antibodies were prepared.The polyclonal antibody and some of monoclonal antibodies could neutralize SLP activity.These results suggest that the antibodies prepared can be referred to SLP target protein.The Western blot assay shows that molecular weight of SLP is about 67 kD,and p I is 4.2.