~（131）I标记肝癌单克隆抗体HAb18F（ab＇）_2的NBS法研究

本文研究了以溴代琥珀酰亚胺（ＮＢＳ）为氧化剂的（１３１）Ⅰ标记肝癌单克隆抗体ＨＡｂ１８Ｆ（ａｂ’）２的方法。探讨了反应体系中ＮＢＳ的用量，Ｆ（ａｈ’）２的用量及Ｎａ（１３１）Ⅰ活度三个因素对标记率的影响，并从反应机理上予以解释。     

抗人活化B淋巴细胞单克隆抗体制备及鉴定

自儿童手术摘除的新鲜扁桃体分离人Ｂ淋巴细胞，经ＰＭＡ（佛波醇）激活后得到人活化Ｂ淋巴细胞，用于免疫ＢＡＢＬ／Ｃ小鼠后，取免疫小鼠之脾细胞与小鼠骨髓瘤细胞ＳＰ２／０融合，融合后细胞于含ＨＡＴ－ＩＭＤＭ完全培养基的２％甲基纤维素半固体培养基选择生长，经细胞酶联免疫吸附（ＣＥＬＩＳＡ）筛选间接免疫荧光流式细胞仪鉴定，获培养上清对活化Ｂ细胞及３Ｄ５细胞株细胞反应阳性，而Ｊｕｒｋａｔ细胞株细胞反应阴必的杂交     

特异识别人脑乙酰胆碱酯酶抗原表位的单克隆抗体1F9

目的 :为了进一步研究乙酰胆碱酯酶的功能和结构以及精确地确定抗体对位与抗原表位的关系 ,我们采用单克隆抗体对人脑AChE的肽库进行筛选 ,希望能找到一对一的表位对位关系。方法 :采用MGeyson的方法化学合成人脑乙酰胆碱酯酶的十肽肽库 ,十肽N端生物素化后用于ELISA检测。结果 :我们用鼠抗电鳐乙酰胆碱酯酶的单抗 1F9筛选人脑乙酰胆碱酯酶的十肽肽库 ,得到 1个抗原表位 111。结论 :抗原表位 111可以被兔抗人脑AChE的多抗和鼠抗电鳐乙酰胆碱酯酶的单抗 1F9识别 ,表明此表位序列在人脑AChE和电鳐AChE中是高度保守的。     

无蛋白培养中杂交瘤细胞的生长与单克隆抗体分泌

目的：建立应用无蛋白培养基大量制备单克隆抗体（McAb)的生产方法。方法：本实验以3株猪瘟病毒特异单抗杂交瘤细胞为研究对象，通过与常规的有血清培养法比较，研究了无蛋白培养基中杂交瘤的细胞生长特性、抗体分泌效率、活性和纯度。结果：无蛋白培养基中单抗的产量与效价比有血清培养的单抗高2－4倍，而且单抗的纯度显著提高。结论：无蛋白培养基可完全取代常规的有血清培养基，用于生产高滴度、高纯度单克隆抗体。     

抗凝血酶受体单克隆抗体的制备与鉴定

采用杂交瘤技术，获得了４株稳定分泌抗凝血酶受体单克隆抗体（ＭｃＡｂ）杂交瘤细胞株。４株ＭｃＡｂ均为ＩｇＧ１κ链。ＥＬＩＳＡ交叉试验结果表明，该ＭｃＡｂ不与人凝血酶、凝血酶原和ＨＣＶ多肽反应。４株杂交瘤细胞培养上清液效价为３．２×１０－２～１．２８×１０－３，腹水效价为１．６×１０－６～５．１２×１０－７。     

促甲状腺激素单克隆抗体的制备和应用

目的:获得高效价、高特异性的促甲状腺激素(TSH)单克隆抗体,为建立高灵敏度的促甲状腺激素水平的测定方法奠定基础。方法:采用细胞融合技术,以TSH抗原免疫BALB/c小鼠的脾细胞与SP2 /0骨髓瘤细胞进行融合。通过多次阳性筛选和有限稀释,建立了抗人TSH单克隆抗体杂交瘤细胞株。并对所建立的单抗进行了鉴定。结果:细胞融合率达100%,阳性率15%。共建立了5株杂交瘤细胞株,均属IgG1 类。且与FSH、HCG、LH无交叉反应。McAb腹水效价在1∶5×105 ~1∶1×107。6个月液氮冻存后复苏培养测定细胞株的稳定性好。结论:本实验的融合率高,阳性率较高,所建立的细胞株的特异性高,McAb腹水效价高。为提高TSH检测的灵敏度和简便性提供了重要条件。     

TLR2和TLR4单克隆抗体对DSS诱导的小鼠结肠炎肠黏膜细胞因子及肠道菌群的影响

Objective To evaluate the effect of TLR2McAb and TLR4McAb on intestinal flora of DSS-induced colitis in mice. Methods Fifty healthy male BALB/c mice (SPF level), were randomly assigned into five groups: the control group( group A), the UC model group( group B), TLR2McAb intervention group( group C), TLR4McAb intervention group( group D) and TLR2McAb + TLR4McAb intervention group(group E). Clinical symptoms were evaluated by the disease activity index(DAI), while tissue sam ples were evaluated by histological scoring(HS). The quantities of mRNA for IFN-γ, IL-4 and IL-17 were determined by real-time PCR. Meanwhile, fecal samples were obtained directly from the cecum for microbiological studies. Results After the treatment with TLR2McAb and TLR4McAb, DAI and HS were decreased significantly. Compared with group A, inflammatory cytokines such as IFN-γ, IL-4 and IL-17 in group B were higher. Compared with group B, expression of these three cytokines in group C to E was all markedly decreased. Group A showed a considerable predominance of Lactobacillus spp and Bifidobacterium spp,while the UC model group showed a conspicuous increase of Escherichia coli and decreases of Lactobacillus spp and Bifidobacterium spp. After treatment with TLR2McAb or/and TLR4McAb, Lactobacillus spp and Bifidobacterium spp increased to the normal level. But counts of E. Coli in the three intervention groups were not changed. Conclusion TLR2McAb and TLR4McAb suppressed the development of DSS-induced colitis and increase cecum counts of Lactobacilli and Bifidobacteria.     

小鼠CTL体外非特异性扩增对其杀瘤活性的影响

目的 在体外用抗CD3单抗、抗CD28单抗和白细胞介素2（IL－2）扩增肿瘤特异性CTL,为肿瘤过继治疗提供足够数量的、具有高度杀瘤活性的效应细胞。方法 采用两种方案培养肿瘤细胞免疫的小鼠脾细胞。（1）抗CD3单抗刺激48h后,加入抗CD3单抗和20U／mlrIL－2（抗－CD3＋IL－2组）;（2）抗CD3单抗和抗CD28单抗同时刺激48h后,加入抗CD3单抗、抗CD28单抗和20U／ml rIL－2（抗－CD3＋抗－CD2＋IL－2组）。分别检测2组效应细胞的增殖水平、杀瘤活性及表型。结果 抗CD3＋IL－2组细胞的^3H－TdR掺入量在第6天、12天、20天分别为22126、52426、2072,抗－CD3＋抗－CD28＋IL－2组细胞的^3H-TdR掺入量在第6天、12天、30天分别为32168、12922、3265,后者明显高于前者。在培养12d时,抗－CD3＋IL－2组细胞对FBL03的最大杀伤活性为66．4％,抗－CD3＋抗－CD28＋IL－2组细胞对FBL－3的最大杀伤活性为77．8％。细胞表型FACS分析结果表明,抗－CD3＋抗－CD28＋IL－2组培养12d后的细胞90％以上为Thy1.2^ 细胞,且CD25^ 细胞在抗－CD3＋抗－CD28＋IL－2组、抗－CD3＋IL－2组分别为23．00％、8．15％。结论 抗CD3单抗、抗CD28单抗和低剂量IL－2同时非特异性刺激,可获得大量扩增的、具有高度杀瘤活性的肿瘤特异性CTL。     

检测血吸虫循环抗原的多价单克隆抗体诊断试剂盒的研制和应用

用杂交瘤技术生产抗血吸虫肠相关循环阳极抗原（ＣＡＡ）单克隆抗体与血吸虫卵糖蛋白单克隆抗体经混合后标记辣根过氧化物酶（ＨＲＰ），制成Ｄｏｔ－ＥＬＩＳＡ诊断试剂盒，检测轻、中、重不同疫区粪检阳性的血吸虫病人血清中循环抗原，阳性率分别为８４．３％、８７．２％和９１．５％。累计阳性率为８９．２％（６１９／６９４）。对健康人、肝吸虫、肺吸虫病人血清均未出现阳性反应，显示有较高的敏感性和特异性。ＥＰＧ＞８０的病人循环抗原检出率高干ＥＰＧ／８０的病人。说明循环抗原检出率与感染度有关。血吸虫病人经吡喹酮治疗后半年和１年循环抗原转阴率分别为７１．４％和８８．６％。结果表明．该试剂盒可用于确诊病人和考核疗效。试剂盒质量稳定，操作简便快速，整个试验可在１小时内完成，且不需特殊仪器，适于现场大规模查病应用。     

PREPARATION AND IDENTIFICATION OF MONOCLONAL ANTIBODIES AGAINST THE EXTRACELLULAR DOMAIN OF INTEGRIN α6 SUBUNIT-THE SPECIFIC LAMININ RECEPTOR

Objective: To prepare monoclonal antibody (McAb) against the Integrin α6 extracellular domain and identify its biological activities. Methods: Fusion-protein of integrin α6 extracellular domain (GST-IAGED) was expressed in E.coli. JM109 and used for immunizing BALB/C mice. The spleen cells from immunized mice were fused with SP2/0 cells and selectively cultured with HAT medium. ELISA and immunocytochemistry staining were used to select hybridomas. Results: One strain of hybridoma cells that secreted specific monoclonal antibody against integrin α6 extracellular domain was indentified. The immunoglobulin subclass of the McAb was IgG1. Conclusion: The McAb against the extracellular domain of integrin α6 was successfully prepared by using GST-IA6ED fusion protein expressed by E.Coli. And the McAb had positive reaction with human hepatocarcinoma cells-BEL-7402.