188Re直接标记抗体方法研究

以2-巯基乙醇作为抗体的还原剂,葡萄糖酸钠作为中间弱配体,在pH5．0的条件下,以SnCl2快速还原”。ReO4^-,进行了单克隆抗体IgG的直接标记。探讨了影响标记率的因素。结果显示,优化后的标记条件为pH5．0,室温下标记3h,SnCl2用量2．66μmol,葡萄糖酸钠用量80μmol。该方法操作步骤简单方便,标记率较高,为94．4％±0．7％,标记物不需分离纯化,且有良好的体外稳定性,室温下可稳定保存8h。     

免疫酸奶的制备

本实验以新鲜免疫常乳为原料，经离收脱脂，采用传统工艺制做既营养又保健的免疫酸乳，研究中发现，免疫酸乳中，IgG含量及活性基本保持不变，保加利亚乳酸杆菌和嗜热链球菌不分解IgG，同时，没有发现免疫常乳中特异性IgG对发酵剂的发酵能力有不良好影响。     

抗黄曲霉毒素B1重链IgG2b亚型单克隆抗体的制备及应用

目的制备抗黄曲霉毒素B1(AFB1)的重链IgG2b亚型单克隆抗体,并建立黄曲霉毒素B1的间接竞争ELISA检测方法。方法采用AFB1-BSA偶联物为免疫抗原,AFB1-STI偶联物为检测抗原,利用间接竞争ELISA筛选分泌抗AFB1单抗的细胞株,并对抗体进行鉴定。结果筛选到1株能分泌特异性抗AFB1单克隆抗体的杂交瘤细胞株,其染色体数目为(90±10)条;所分泌的单抗亚类重链为IgG2b,轻链为λ;间接ELISA检测该株杂交瘤细胞分泌上清效价在1∶12800～1∶25600,纯化腹水效价为1∶105～1∶106。传30代及液氮中保存6个月,抗体效价稳定;建立的间接竞争ELISA检测方法的最低检出限为0.001ng/ml,校正曲线的线性范围为0.005～5ng/ml,IC50为0.01ng/ml;与AFB1的结构类似物AFM1和化学结构有差异的脱氧雪腐镰刀菌烯醇的交叉反应率分别为1.4%和<1%。结论所制备的抗黄曲霉毒素B1单克隆抗体具有高度特异性和稳定性,可应用于间接竞争ELISA检测方法。     

STAg联合蜂胶佐剂鼻内免疫小鼠抗弓形虫感染的保护作用

目的探讨可溶性速殖子抗原(STAg)联合蜂胶佐剂鼻内免疫小鼠抗弓形虫感染的保护作用及蜂胶的适宜剂量。方法将60只BALB/c小鼠随机分为5组,每组12只,分别用20μgSTAg、20μgSTAg+5μg蜂胶、20μgSTAg+10μg蜂胶、20μgSTAg+20μg蜂胶及20μgSTAg+40μg蜂胶鼻内免疫小鼠2次,间隔14d。末次免疫后第10天,用RH株弓形虫速殖子灌胃攻击,4×104个/只,观察小鼠存活情况。攻击后第30天处死全部小鼠,分离计数小鼠脑、肝组织内速殖子和肠系膜淋巴结淋巴细胞(MLNL)、脾T淋巴细胞;ELISA法检测小鼠肺冲洗液、粪便弓形虫特异性sIgA和血清IgG抗体水平。结果随着蜂胶剂量的增加,小鼠脑、肝速殖子数呈下降趋势,20μgSTAg+40μg蜂胶组显著低于20μgSTAg组,脑、肝组织减虫率分别为46.10%和60.11%;MLNL和脾T淋巴细胞增殖活性提高,其中20μgSTAg+40μg蜂胶组小鼠MLNL和脾T淋巴细胞数分别为(2.09±0.24)×107和(1.89±0.21)×107,均显著高于20μgSTAg组;肺冲洗液、粪便弓形虫特异性sIgA和血清IgG抗体水平呈增高趋势,但各组间差异无统计学意义。结论STAg联合蜂胶佐剂鼻内免疫小鼠可有效诱导抗弓形虫感染的保护作用,适宜的蜂胶剂量为40μg。     

Probing Specific Interaction Forces Between Human IgG and Rat Anti-Human IgG by Self-Assembled Monolayer and Atomic Force Microscopy

Interaction forces between biological molecules such as antigen and antibody play important roles in many biological processes, but probing these forces remains technically challenging. Here, we investigated the specific interaction and unbinding forces between human IgG and rat anti-human IgG using self assembled monolayer (SAM) method for sample preparation and atomic force microscopy (AFM) for interaction force measurement. The specific interaction force between human IgG and rat anti-human IgG was found to be 0.6–1.0 nN, and the force required for unbinding a single pair of human IgG and rat anti-human IgG was calculated to be 144 ± 11 pN. The results are consistent with those reported in the literatures. Therefore, SAM for sample preparation combined with AFM for interaction measurement is a relatively simple, sensitive and reliable technique to probe specific interactions between biological molecules such as antigen and antibody.     

Sera from Patients with NMOSD Reduce the Differentiation Capacity of Precursor Cells in the Central Nervous System

Introduction: AQP4 (aquaporin-4)–immunoglobulin G (IgG)-mediated neuromyelitis optica spectrum disorder (NMOSD) is an inflammatory demyelinating disease that affects the central nervous system, particularly the spinal cord and optic nerve; remyelination capacity in neuromyelitis optica is yet to be determined, as is the role of AQP4–IgG in cell differentiation. Material and Methods: We included three groups—a group of patients with AQP4–IgG-positive neuromyelitis optica, a healthy group, and a sham group. We analyzed differentiation capacity in cultures of neurospheres from the subventricular zone of mice by adding serum at two different times: early and advanced stages of differentiation. We also analyzed differentiation into different cell lines. Results and Conclusions: The effect of sera from patients with NMOSD on precursor cells differs according to the degree of differentiation, and probably affects oligodendrocyte progenitor cells from NG2 cells to a lesser extent than cells from the subventricular zone; however, the resulting oligodendrocytes may be compromised in terms of maturation and possibly limited in their ability to generate myelin. Furthermore, these cells decrease in number with age. It is very unlikely that the use of drugs favoring the migration and differentiation of oligodendrocyte progenitor cells in multiple sclerosis would be effective in the context of neuromyelitis optica, but cell therapy with oligodendrocyte progenitor cells seems to be a potential alternative.     

共轭亚油酸对小白鼠身体组成的影响

饲料中添加不同剂量共轭亚油酸喂养 4周龄小白鼠 4周。实验结果表明 ,血清中Chol、HDL和LDL含量的平均值随着共轭亚油酸添加量的增加而升高 ,而Chol/HDL的比值则几乎保持不变。饲料中添加 0 .5 %的共轭亚油酸对血清中甘三酯浓度影响不大 ,基本上类似于对照组的含量 ;当共轭亚油酸的添加量达到 1%时 ,与对照组间差异较大。共轭亚油酸对血清中免疫球蛋白含量影响不大。小白鼠身体水分和蛋白质含量分析结果表明 ,饲料中添加共轭亚油酸增加了水分和蛋白质含量。此外 ,共轭亚油酸能显著降低小白鼠身体的脂肪含量 ,这主要通过抑制LPL活性和增加肉碱棕榈酰转移酶活性来实现 ;同时通过抑制肝脏中硬脂酰辅酶A去饱和酶mRNA的表达来改变其脂肪酸组成。     

羊抗人IgG和出血热单抗(E_5)在辐射改性的硅橡胶膜上固定化研究

本文用γ辐射技术,在硅橡胶膜(Silastic)上接枝丙烯酰胺单体(AAM),用不同接枝率的Silastic-AAM膜通过叠氮化后对羊抗人IgG和出血热单克隆抗体(E_5)进行了偶联固定化。同位素~(125)碘标记方法对固定在Silastic-AAM膜上的羊抗人IgG量与接枝率的关系,以及抗体量与其免疫活性的关系进行了测定,得到了适于固定化的抗体偶联反应液最适浓度和最佳接枝率范围;用ELISA法对Silastic-AAM膜固定化的出血热(E_5)单抗进行了免疫活性实验,得到了承某一实验条件下具有明显的阳性反应。结果表明,经适当改进,本法对临床诊断有一定应用价值。     

IgG与HA对TZNT合金腐蚀行为的影响

在PBS溶液(pH=7.4)中采用电化学测试技术考察了免疫球蛋白G(IgG)与透明质酸(HA)对潜在的人工关节用TZNT合金腐蚀行为的影响。结果表明:免疫球蛋白G的吸附可在一定程度上减缓TZNT合金表面氧化膜的生长;透明质酸抑制TZNT合金腐蚀的效果与其掺入浓度和浸泡时间相关,在其浓度为2.5 g/L,浸泡72 h时,抑制合金腐蚀的效果最佳。