改良火箭免疫电泳法测定静脉注射用丙种球蛋白的IgG含量

目的　建立一种特异性的检测静脉注射用人免疫球蛋白中的 Ig G含量的方法。方法　用火箭免疫电泳法 ,以不同浓度的甲醛对 Ig G甲酰化处理 ,再进行免疫电泳。用 Ig G标准品建立标准曲线 ,测定样品中的Ig G含量。结果　用该法制备的标准曲线有良好线性关系 (r=0 .996 7)。对 3批样品做重复性测定 (n=6 ) ,其百分含量分别为 5 .37%± 0 .2 1% ,5 .2 8%± 0 .39%和 5 .80 %± 0 .30 % ,重复性好。结论　该法可以作为血液制品的常规检测方法     

Isolation and partial characterization of two distinct types of antithrombin III from rabbit

Heparin-Sepharose chromatography of rabbit plasma or serum yields two fractions of free antithrombin III (AT). The first of these elutes at a lower salt concentration, represents about 90% of the AT in plasma, and has an approximately 2000 dalton higher molecular weight by SDS polyacrylamide electrophoresis. An antibody to the lower affinity species reacts with the second form. The higher affinity AT is not formed from the lower affinity type during blood coagulation as demonstrated by approximately equal levels in plasma and serum, and lack of conversion of ¹²⁵I-labelled lower affinity AT to the higher affinity form during blood coagulation. Heparin cofactor and progressive AT activities of the two forms are essentially identical when assayed by chromogenic substrates. The forms are separable by crossed immunoelectrophoresis.     

Immunoelectrophoresis in the diagnosis of neuroectodermal and mesodermal intracranial tumours,especially those of the posterior fossa

Summary Immunoelectrophoresis of extracts of 200 intracranial tumours against rabbit anti-glioblastoma serum gave positive results (= precipitation) in all cases of tumours of neuroectodermal origin such as glioblastoma, astrocytoma, oligodendroglioma, ependymoma, neurinoma, and spongioblastoma. No immunoelectrophoretic precipitation was seen for any of the tumours of mesenchymal origin, for instance meningioma and metastases of cancer. On the basis of these findings, immunoelectrophoresis is considered to be a reliable method for differentiation between tumour tissue of neuroectodermal and non-neuroectodermal origin.Among the 41 posterior fossa tumours some unusual observations were made. Cerebellar angioblastoma (Lindau tumour) showed an atypically located precipitation line, which for the present is interpreted as an immunological reaction to vascular wall tissue. Furthermore, among the group of so-called medulloblastomas, two subgroups were distinguished on the basis of three parameters. The first of these subgroups comprises tumours whose immunoelectrophoretic pattern resembles that of gliomas, which are histologically characterized by neuroectodermal structures and which occur in younger children (5–10 years). The tumours of the second subgroup, which do not show this neuroectodermal immunoelectrophoretic pattern, have a sarcomatous character histologically, and occur in patients aged between 10 and 50 years. The view that medulloblastoma comprises a number of different types of tumour seems to be confirmed by this finding.     

火箭免疫电泳法定量检测4价脑膜炎球菌多糖疫苗中C群脑膜炎球菌多糖含量的方法学验证

目的 验证用火箭免疫电泳法(rocket immunoelectrophoresis,R1E)检测4价脑膜炎球菌多糖疫苗(tetravalent serogroup A／C／W135／Y meningDcoccal polysaccharide vaccine,MPV4)中C群多糖含量的可靠性。方法 以系列浓度的C群多糖溶液为标准,采用RlE对MPV4中C群多糖含量迸行重复测定。结果 最佳线性范围为30～86 mg／L,相关系数(r)均大于0.985;MPV4与C群多糖参比品的剂量反应曲线之间具有良好的平行性;在精密度试验中,试验内CV为6.08％～8.07％,试验间Cy为7.24％～9.19％;A、W135和Y群多糖及乳糖不引起非特异性免疫反应。结论 本法的线性、精密度和专属性均符合验证要求,可作为定量检测MPV4中C群多糖含量的方法。     

A、C、Y、W135群脑膜炎球菌多糖疫苗多糖含量测定方法的建立

目的  建立测定A、C、Y、W135群脑膜炎球菌（meningococcus,Men）多糖疫苗（groups A,C,Y,W135 menignococcal polysaccharide vaccine,MPV4）多糖含量的火箭免疫电泳（rocket immunoelectrophoresis,RIE)法。方法  分别用A、C、Y、W135群Men菌液、A、C、Y、W135群Men菌液加佐剂、A、C、Y、W135群Men多糖-牛白蛋白结合物免疫家兔,制备特异性抗血清。将制备的抗血清以一定的比例加入琼脂糖凝胶制成平板,并将纯化的多糖作为定量参考品加入抗原孔进行RIE。以多糖含量和对应的RIE峰高作标准曲线并建立直线回归方程。采用优化的RIE法测定MPV4多糖含量和分子大小。结果  菌液加佐剂制备的抗血清效价较理想。在确定的电泳条件下,建立的RIE法制备的标准曲线呈现良好的线性关系,相关系数值均大于0.98。该法特异性较好,未检出各多糖间的交叉反应。采用该法测定3批MPV4多糖含量、分子大小及回收率的结果均与先前的检定结果相符,均符合质控标准。结论  建立的RIE法可作为MPV4多糖抗原含量的测定方法。     

Effects of iodipamide on human C3 and factor B in vitro

The effects of iodipamide on C3 and factor B in normal human serum and in purified form have been examined by immunoelectrophoresis and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Temperature-dependent changes in immunoelectrophoretic profile's have been observed; however, these are not the same as those obtained after treatment of normal human serum (NHS) with cobra venom factor Naja naja. Analyses of iodipamide-treated NHS and purified C3 and factor B by reducing SDS-PAGE indicate that no macromolecular changes have occurred in C3 and factor B that can be ascribed to proteolysis (i.e., activation). The changes observed in C3 and factor B, including loss of hemolytic activity, appear to be due to direct interactions between iodipamide and C3 and factor B. In the case of factor B, iodipamide treatment at 37° C induces aggregation, which is reversible upon reduction with β-mercaptoethanol.     

Characterization of an antigen in acute anaphylactic dialysis reactions: ethylene oxide-altered human serum albumin

Anaphylactic reactions to dialysis have been associated with antibody to human serum albumin (HSA) that has been reacted with ethylene oxide (ETO). ETO, an agent used to sterilize dialyzers, is a potent alkylator of proteins. We have studied the antigenic characteristics of ETO-HSA. By use of immunoelectrophoresis we have demonstrated differences in electrophoretic mobility between HSA and ETO-HSA. By gel chromatography we have observed an apparent increase in the molecular weight of ETO-HSA as compared to HSA. Antigenic activity of ETO-HSA as determined by the technique of Lidd and Farr is found in all molecular weight fractions. Cutaneous reactivity to ETO-HSA can be transferred to subhuman primates. That reactivity is lost if the sera are heated to 56 degrees C for 4 hours. Antibody activity to ETO-HSA cannot be inhibited by ETO-glycine or HSA but can partially be inhibited by ETO-polylysine or ETO-ovalbumin. This suggests that antibody is directed against a new antigenic determinant on the ETO-protein moiety, probably an ETO-amino acid determinant like lysine.