单克隆抗体与葡萄球菌肠毒素A结合物的制备及其抗肝癌作用

目的：制备超抗原（BAg)葡萄球菌肠毒素A（SEA）与抗人原发性肝细胞癌（简称肝癌）单克隆抗体（McAb)HAb18F(ab‘)2段的结合物,探讨其介导外周血单个核细胞（PBMC）的抗人肝癌作用。方法,制备McAb HAb18,并用木瓜蛋白酶消化法制备其F（ab‘)2段,再用化学连接剂N－琥珀酰亚氨基－3－（2－二硫吡啶）丙酸酯（SPDP）制备HAb18 F(ab‘)2-SEA结合物,结合物经经快速蛋白质液相层析系统用凝胶色谱柱纯化后,用SDSPAGE鉴定各种收集峰,免疫组化ABC法鉴定结合物的抗体活性,MTT法鉴定结合物活化PBMC的活性及其介导PBMC的杀伤肝癌细胞作用,结果,制备的HAb18F(ab‘)2-SEA结合物具有肝癌抗体活性和活化PBMC的功能,此结合物具有明显的介导PBMC杀伤肝癌细胞的作用,并与其[浓度呈正相关,结论,SPDP是一种很好的蛋白质交联剂,HAb18 F(ab‘)2-SEA结合物民向治疗肝癌具有一定的发展前景。     

Anti-DNA antibodies: aspects of structure and pathogenicity

Anti-DNA antibodies contribute to the pathology of systemic lupus erythematosus. Their depositon in tissue lesions could result from localization of preformed immune complexes of antibodies with DNA or nucleosomes, or from cross-reaction of anti-DNA antibodies directly with tissue proteins. Structural analyses contribute to understanding their pathogenic potential. Primary structures of lupus immunoglobulin G double-stranded DNA-binding autoantibodies are determined by immunoglobulin genes with mutated variable region segments, indicative of selection by immunizing antigen. Arginine, lysine and asparagine residues in complementarity-determining region favor DNA binding. Heavy-chain variable regions make major contributions to DNA binding; affinity and specificity of binding are modulated or can be abrogated by the light-chain variable domain. Crytallographic structure is known for a few antibody-DNA complexes and several ligand-free Fab fragments. Computer modeling supplements this limited information. Structural information of lupus antibody interactions with both DNA and cross-reacting molecules will support use of ligands to inhibit tissue deposition of the antibodies and prevent lesion formation in lupus. Received 4 July 2002; accepted 23 July 2002 RID="*" ID="*"Corresponding author.     

丙型肝炎病毒抗体免疫酶快速检测试剂的评价

对丙型肝炎病毒抗体免疫酶快速检测试剂(IEA)检测丙型肝炎病毒抗体(HCV-Ab)的性能、结果等技术参数进行评价,为实验室的选用提供参考.采用IEA法、ELISA法和胶体金法对中国生物制品检定所“第三代ELISAHCV-Ab诊断试剂国家参考品”和3297份血清或血浆标本同时测定,对IEA的敏感性、特异性、重复性和检测时间等结果进行评价.结果表明:IEA可通过中国生物制品检定所“第三代ELISAHCV-Ab诊断试剂国家参考品”检验;与ELISA法比较,IEA的检出率为99.06%,灵敏度与ELISA相当,胶体金的检出率为86.01%,灵敏度比ELISA低;IEA检测所需时间15～25min.IEA既有ELISA的敏感性、特异性,又有胶体金简便、快速的特点.适用于献血员及高危人群中HCV抗体的筛查和临床诊断HCV感染,以及门诊、急诊和基层医疗单位的现场检测.     

基因工程菌株产生人类绒毛膜促性腺激素抗原的研究——Ⅰ.β-Galactosidase-hCG的分离纯化与鉴定

通过寡核苷酸合成仪,用化学方法合成相当于hcGβ链C末端36肽基因的片段,成功地克隆到噬菌体载体λgt11中,并在宿主菌株E. coli y_(1089)中得到表达。含λgt11 hCG重组体的E. coli y_(1089)所合成的含人类绒毛膜促性腺激素β链c末36肽的杂交蛋白(下称:β—galactosidase—hCG),通过硫酸铵沉淀、亲和层析和制备性聚丙烯酰胺凝胶电泳,可得电泳纯单一色带的β—gaIactosidase—hCG。蛋白质转印(Western Blot)技术证实β—galactasidase—hCG具有hCGβ链c末端36肽的免疫特性。对流免疫电泳和免疫双扩散试验结果表明β—galactosidase—hCG具免疫原性。从一立升培养液的菌体中可分离纯化约3.8毫克的β—galaetosidase—hCG。     

Co-localization of glucagon and pancreatic polypeptide in testudine pancreas

Summary Immunocytochemistry was carried out on sections of pancreas from the gopher tortoise,Gopherus polyphemus. Combined immunofluorescent and peroxidase-anti-peroxidase techniques showed unequivocally that some of the cells were immunoreactive for both glucagon and pancreatic polypeptide (PP). Antibodies directed against avian PP, bovine PP, and human PP all have a positive reaction. Co-localization of glucagon and PP in the pancreas of the gopher tortoise indicates that the occurrence of these hormones in the same cells is widespread in higher vertebrates.Acknowledgments. This work was supported in part by grants from the University of Cape Town, the M. Crossley bequest, the N. Atkinson bequest, and Herman and Caporn bequests. The generous gifts of antisera by Dr Chance and the late Dr Kimmel, and the gift of hormones by the Lilly Research Laboratories are gratefully acknowledged. The excellent technical assistance of J. Thompson and J. B. De Haan is also gratefully acknowledged. Please send all correspondence to the USA address.