摘要
应用淋巴细胞杂交瘤技术获得１０株抗重组人肿瘤坏死因子－α（ｒＨｕＴＮＦ－α）ＭｃＡｂ。７株ＭｃＡｂ为ＩｇＧ１，２株为ＩｇＧ２ａ，１株为ＩｇＧ２ａ。１０株ＭｃＡｂ与淋巴毒素（ＬＴ或ｒＨｕＴＮＦ－β）、ｒＨｕＩＬ－６和载体菌菌体蛋白均无交叉反应。其中７株ＭｃＡｂ具有中和活性，７株ＭｃＡｂ能识到天然的ＴＮＦ－α。Ｇ１１和Ｅ６ＭｃＡｂ应用ＡＢＣ法免疫组化染色肝病患者组织细胞片得到明确的阳性结果。用２株识别不同表位的ＭｃＡｂ建立了一步夹心ＥＬＩＳＡ法，检测ＴＮＦ－α的敏感性可达到１００ｐｇ／ｍｌ左右。
Ten hybtidoma cell lines were preparated by the lymphocyte hybridoma technique. Of theten McAbe,seven(B5,D12,E6,E11,F6,F8,G9)were IgG1,two(C4,D2)were IgG2band one(G11)was IgG2a.None of the McAbs cross-reacted with rHuTNF-β,rHuIL-6 or normal E.coli protein.Seven McAbs(B5,C4,D2,E6,F6,F8,G9)could inhibit the binding of rHuTNFαto it's receptor.Seven McAbs(C4,D2,D12,E6,E11,F6,G11)reacted with natural TNFα.The positive staining was observed on hepatocytes of the patients with chronic persistent hepatitis(CPH),chronic active hepatitis(CAH)and liver cirrhoris(LC)by ABC immunohistochcmical methodwith rHuTNF-α McAb G11 and E6.One step sandwich ELISA for detecting TNF-αwas established byusing two McAbs reacted with different epitopes of rHuTNF-α,and the sensitivity of the method is approximately 100 pg/ml.
