前列腺特异性抗原免疫放射分析反应的动力学研究

本文着重对前列腺特异抗原（ＰＳＡ）的免疫放射分析的反应时间进行研究。实验采用两步，一为固定第一步时间，发迹第二步反应时间，另一固定第二步时间，改变第一步反应时间，观察反应的变化。结果表明，第一反反应０．５小时以后，标准和质控的ｃｐｍ已相对恒定，０．５小时的最大结合率为５８．７％，与６小时的最大结合率５５．８％相比，差异不显著。第二步反应，从１小时到６小时，结合率随时间的增加而升高。１小时的结合率为     

前列腺特异抗原免疫放射分析反应的动力学研究

本文着重对前列腺特异抗原（ＰＳＡ）的免疫放射分析的反应时间进行研究。实验采用两步，一为固定第一步时间，改变第二步反应时间，另一为固定第二步时间，改变第一步反应时间，观察反应的变化。结果表明，第一步反应０．５小时以后，标准和质控的ｃｐｍ已相对恒定，０．５小时的最大结合率为５８．７％，与６小时的最大结合率５５．８％相比，差异不显著。第二步反应，从１小时到６小时，结合率随时间的增加而升高。１小时的结合率为４６．３％，与６小时的结合率７２．２％相比，差异显著。此结果将有助于对ＰＳＡ的临床检测提供一个最适宜的反应时间。     

抗人血浆纤维结合蛋白单克隆抗体的初步研究

本文应用人血浆纤维结合蛋白免疫ＢＡＬＢ／ｃ小鼠，经细胞融合制备出一组（５株）人血浆纤维结合蛋白的单克隆抗体。经免疫酶标和免疫组化检测，这组单抗仅特异地与人血浆纤维结合蛋白结合，而与血浆中的其它大分子糖蛋白如纤维蛋白原、凝血酶敏感蛋白和胶原等均无明显的交叉反应。与多种器官，组织的间质可发生性异性反应，而与组织中的细胞则无反应，间接免疫荧光测定，这组抗体与正常人的造血细胞及某些造血细胞系和部分白血病病     

多反应性单克隆抗体的免疫生化分析

探讨多反应性抗体的交叉反应机制是免疫学基础理论研究的重要课题。我们曾研制了一批多反应性单克隆抗体，其中抗体ＸＹ１２不仅能够与中间纤维蛋白结合，而且还与ＤＮＡ发生反应。本文采用免疫生化的方法，产生、纯化并分析了抗体ＸＹ１２的Ｆａｂ片段。ＥＬＩＳＡ和放射免疫印迹结果表明，这些Ｆａｂ片段仍具有完整抗体的多反应性。竞争性抑制实验显示，一定量的波形蛋白能够有效地抑制抗体ＸＹ１２与ＤＮＡ的结合。这些结果提示抗体ＸＹ１２的对两种抗原的反应确实都发生在抗体与抗原的结合区域而非抗体的其它部位。     

聚苯乙烯—孕酮免疫微球的新法制备与检测

本研究采用改进的无乳化剂乳液聚合法制备聚苯乙烯微球，并对聚合反应温度和有机溶剂含量对聚合反应和粒径的影响进行了研究；再将合成的聚苯乙烯微球与孕酮抗体反应，结果表明通过加入少量有机溶剂，提高了聚合反应速度和转化率，制备出了粒径可控的单分散聚苯乙烯免疫微球，粒径在２００～８００ｎｍ之间，微球具有较高的抗体结合容量，且结合后保持了较高的抗体活性。用合成的孕酮免疫胶乳进行免疫凝集实验，观察了反应时间，反应温度的影响，确定了免疫反应的基本反应条件。     

抗人胎胰相关抗原单克隆抗体C1免疫组化研究

用人胎胰提取物免疫ＢＡＬＢ／ｃ小鼠，获得能稳定分泌ＩｇＧ２ａ单抗的杂交瘤细胞Ｃ１。免疫组化染色表明，Ｃ１与胎胰、胰腺癌、胃癌和大肠癌等组织有较强结合反应，与正常胰腺、十二指肠、胃、大肠等处的消化腺上皮细胞有轻度结合反应。初步表明，通过定量分析单抗，Ｃ１对消化道癌肿的诊断有一定参考价值。     

抗人胎胰相关抗原单克隆抗体C1免疫组化研究

用人胎胰提取物免疫ＢＡＬＢ／ｃ小鼠，获得能稳定分泌ＩｇＧ２ａ单抗的杂交瘤细胞Ｃ１。免疫组化染色表明，Ｃ１与胎胰、胰腺癌、胃癌和大肠癌等组织有较强结合反应，与正常胰腺、十二脂肠、胃、大肠等处的消化腺上皮细胞有轻度结合反应。初步表明，通过定量分析单抗，Ｃ１对消化道癌肿的诊断有一定的参考价值。     

两种亚型ADHD儿童的反应抑制

目的：检验两类ADHD儿童（注意缺陷型和混合型）在两种反应抑制功能-反应冲突和反应停止上的表现。方法：采用Stroop任务和Go/NoGo任务的结合，在计算机上逐一呈现每个试验任务。要求实验儿童按指导语对刺激作按键或不按键的反应，计算机记录下儿童的反应时和错误率。结果：在反应冲突控制能力上，未发现ADHD儿童总体上正常对照组儿童有明显差别。在反应停止能力上，ADHD儿童明显弱于正常对照组儿童，这种弱势主要来自混合型ADHD儿童。结论：上述结果提示，ADHD儿童在反应冲突和反应停止上的缺损程度不同。两类ADHD儿童的认知和神经机制方面的缺陷也可能不同。     

p53基因结构与功能研究的新进展

ｐ５３蛋白的信号区有１个ＳＨ－３区结合部位，非专一ＤＮＡ结合区可促进顺序的ＤＮＡ结合区与ＤＮＡ损伤部位结合。Ｐ５３蛋白可整合细胞对各种危急情况的反应，其信息传递途径上游是各种危急状况，有ＡＴＭ类的基因起作用。     

一氧化氮合酶与乙酰胆碱酯酶在大鼠心内神经节细胞共存的组化研究

用还原型尼克酰胺腺嘌呤二核苷磷酸黄递酶反应，结合乙酰碱酯酶组化技术，观察了Ｗｉｓｔａｒ大鼠房后壁内神经节，发现节内存在还原型尼克酰胺泉嘌呤二核苷磷酸黄递酶反应阳性细胞；少量细胞极度深度，并在深染细胞中发现有双突起的细胞：多数轻、中度着色的细胞浆内有蓝色颗粒和棕色反应。     

Effects of addition of salt to bread on IgE antibody responses

The effect of the addition of NaCl to bread on antigen-antibody reaction involving IgE was examined. We compared the antigen-antibody reaction involving IgE between dough including NaCl and that not including NaCl. Crude proteins extracted from the dough including NaCl showed weaker antigen-antibody reactions than proteins from the dough no added NaCl on allergic tests such as precipitin ring test with human-specific IgE, and the IgE binding activity on ELISA. The crude proteins extracted from the dough with NaCl or without NaCl were applied to an affinity chromatography column of immobilized-trypsin chitin. Then proteins having affinity were recovered. The recovered proteins were separated by SDS-PAGE. Each protein was examined for the IgE binding activity on ELISA. When the amount of protein in dough including NaCl was compared with that in dough without NaCl, a significant difference in the amount of ovomucoid between the two samples were confirmed. The IgE binding activity of baked bread was studied as well. Crude proteins of baked bread made from dough including NaCl showed weaker IgE binding activity on ELISA than proteins of baked bread made from dough without NaCl. The importance of NaCl as an ingredient of baked bread was confirmed.     

Effects of addition of salt to bread on IgE antibody responses

The effect of the addition of NaCl to bread on antigen-antibody reaction involving IgE was examined. We compared the antigen-antibody reaction involving IgE between dough including NaCl and that not including NaCl. Crude proteins extracted from the dough including NaCl showed weaker antigen-antibody reactions than proteins from the dough no added NaCl on allergic tests such as precipitin ring test with human-specific IgE, and the IgE binding activity on ELISA. The crude proteins extracted from the dough with NaCl or without NaCl were applied to an affinity chromatography column of immobilized-trypsin chitin. Then proteins having affinity were recovered. The recovered proteins were separated by SDS-PAGE. Each protein was examined for the IgE binding activity on ELISA. When the amount of protein in dough including NaCl was compared with that in dough without NaCl, a significant difference in the amount of ovomucoid between the two samples were confirmed. The IgE binding activity of baked bread was studied as well. Crude proteins of baked bread made from dough including NaCl showed weaker IgE binding activity on ELISA than proteins of baked bread made from dough without NaCl. The importance of NaCl as an ingredient of baked bread was confirmed.     

The effect of excess antigen on the enzyme-linked immunosorbent assay of onco-placental proteins

The time course of the antigen-antibody reaction between human chorionic gonadotrophin, pregnancy specific β₁ glycoprotein and pregnancy associated plasma protein A and their respective immobilised antibodies has been investigated. All 3 antigens showed enhanced binding with increasing length of incubation up to a maximum value, followed by a decrease. It is suggested that the diminution in binding at high antigen concentration is caused by the solubilisation of the immobilised antigen-antibody complex and that this is promoted by the high concentration of antigen.     

Determination by ellipsometry of the affinity of monoclonal antibodies

The reaction between monoclonal antibodies and surface-immobilised hapten was studied by ellipsometry, a method allowing absolute measurement of the surface concentration of proteins. Monoclonal antibodies against 2-phenyloxazolone were used and their affinity for the antigen in solution was determined by calculations of the equilibrium constant from data obtained by measuring fluorescence quenching of the hapten due to antibody binding. The binding rate of antibody to surface-immobilised hapten and the dissociation rate of the complex were measured by ellipsometry. The equilibrium constant of the heterogeneous antigen-antibody reaction was determined by a Scatchard plot. The affinity of the antibodies for the antigen was found to be higher in the heterogeneous than in the homogeneous reaction by a factor which varied between different monoclonal antibodies.     

Cell labeling and sorting by rosette formation. Use of biotin-avidin binding

The binding of biotin by avidin has proven to be a useful adjunct to antigen-antibody reactions in a number of immunological labeling systems. This work has applied it to rosette formation, for identification and separation of selected lymphocytes. Three procedures were compared which incorporated the biotin-avidin reaction into a model system for the rosetting of murine T lymphocytes. The reaction was most effective in conjunction with antibodies binding to the red cell, instead of using red cells which were biotinated on endogenous cell surface proteins. Physical separation of these rosettes from unrosetted cells was demonstrated with magnetic filtration, in a preliminary experiment. One interpretation of differences in effectiveness of rosette formation in the three systems tested is in terms of the flexibility of the antibody molecule, as compared with avidin. The principal finding was that, if employed in a multiple-stage 'sandwich' labeling system, avidinbiotin binding was fully effective and provided new versatility in the design of such rosetting schemes.     

Design and Simulation of Active Biochip System

To solve the problems existing in passive biochip systems, we designed a novel active biochip system. This system introduces negative pressure and controlling devices to adjust the antigen-antibody reaction on the nitrocellulose membrane. Computational simulation demonstrated that this system is a rapid, stable, robust and practical system that may enhance the efficiency of antigen-antibody reactions and improve the repeatability and accuracy of biochip analysis.     

Protective and immunochemical activities of monoclonal antibodies reactive with the Bacillus anthracis polypeptide capsule

Bacillus anthracis is surrounded by a polypeptide capsule composed of poly-gamma-d-glutamic acid (gammaDPGA). In a previous study, we reported that a monoclonal antibody (MAb F26G3) reactive with the capsular polypeptide is protective in a murine model of pulmonary anthrax. The present study examined a library of six MAbs generated from mice immunized with gammaDPGA. Evaluation of MAb binding to the capsule by a capsular "quellung" type reaction showed a striking diversity in capsular effects. Most MAbs produced a rim type reaction that was characterized by a sharp increase followed directly by a decrease in refractive index at the capsular edge. Some MAbs produced a second capsular reaction well beneath the capsular edge, suggesting complexity in capsular architecture. Binding of MAbs to soluble gammaDPGA was assessed by a fluorescence perturbation assay in which a change in the MAb intrinsic fluorescence produced by ligand binding was used as a reporter for antigen-antibody interaction. The MAbs differed considerably in the complexity of the binding curves. MAbs producing rim type capsule reactions typically produced the more complex binding isotherms. Finally, the protective activity of the MAbs was compared in a murine model of pulmonary anthrax. One MAb was markedly less protective than the remaining five MAbs. Characteristics of the more protective MAbs included a relatively high affinity, an immunoglobulin G3 isotype, and a complex binding isotherm in the fluorescence perturbation assay. Given the relatively monotonous structure of gammaDPGA, the results demonstrate a striking diversity in the antigen binding behavior of gammaDPGA antibodies.     

Cell Membrane Antigen-Antibody Complex Dissociation by the Widely Used Glycine-Hcl Method: An Unreliable Procedure for Studying Antibody Internalization

Methods following the process of binding and internalization of antibodies to cell surface antigens have often employed low pH isoosmolar buffers in order to dissociate surface antigen-antibody complexes. One of the most widely used buffers is a 0.05 M glycine-HCL buffer pH 2.8. Since the efficacy of action of this buffer was critical to a series of internalization experiments employing monoclonal antibodies (Mabs) to carcinoembryonic antigen (CEA) expressing cancer cell lines in this laboratory, we tested its performance in a number of different assays. Our results indicate that this buffer only partially dissociates antigen-antibody bonds and therefore can introduce major inaccuracies in internalization experiments.     

固相表面的抗原抗体反应模型与主动式生物芯片系统

为了克服现有被动式生物芯片的不足，我们从抗原抗体反应的基本原理出发，建立了固相表面的抗原抗体反应模型．并根据模型设计了一种新型的主动式蛋白芯片系统，该系统中引入负压发生装置及控制装置，可对硝酸纤维索（NC）膜上发生的抗原抗体反应进行控制。并对该系统进行了仿真，结果表明它具有快速、稳定、鲁棒性好等优点，能满足实际要求。该系统将有助于提高抗原抗体反应的效率，改善芯片检测结果的重复性和准确性。     

Measurement of antigen-antibody complexes in mouse sera by conglutinin, C1q and rheumatoid factor solid phase binding assays

The application of three solid phase tests for the detection of antigen-antibody complexes in the sera of inbred and selectively-bred mice with experimentally induced chronic antigen-antibody complex disease is described. Two of the tests used--the C1q binding assay (C1qBA) and the conglutinin binding assay (KBA)--were previously described methods which we have modified for the detection of murine complexes. The third test is a new solid phase polyclonal rheumatoid factor binding assay (RFBA). The results demonstrate that the correlation of positivity with the three methods varied during the course of a chronic disease but in general results with the KBA showed a better correlation with the RFBA than with the C1qBA. Furthermore, the KBA and RFBA preferentially bound complexes of lower molecular weight than did the C1qBA. It is suggested that the parallel use of these three tests during the study of murine antigen-antibody complex disease will provide valuable information concerning the nature of the complexes involved; since the three tests, when run in parallel, will detect large complement fixing complexes (C1q), small complement fixing complexes (KBA) and small non-complement fixing complexes (RFBA).