首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 62 毫秒
1.
金葡菌C1型肠毒素免疫BALB/C小鼠,取其脾细胞与Sp2/0小鼠骨髓细胞融合选出了能分泌高滴度McAb的18个克隆株,对其中10株进行了鉴定,6档属IgG1,4株属IgG3。用双抗体夹心法和特异性中和抑制试验比较了McAb和PcAb的敏感性的特异检出1ng SEC1/0.5g食物/ml。  相似文献   

2.
人心肌肌钙蛋白T单克隆抗体的研制及鉴定   总被引:1,自引:0,他引:1  
以人心肌肌钙蛋白T(cTnT)为抗原,采用脾内免疫法,免疫BALB/c小鼠,取其脾淋巴细胞与小鼠Sp2/0细胞融合,经间接ELISA法筛选,三次克隆化后获得5株能稳定分泌抗cTnT单克隆抗体(McAb)的杂交瘤细胞G3、G8、G10、A5和A7。免疫球蛋白亚类鉴定其中1株为IgG2a,4株为IgM。染色体数目92~110条。将G3、G8、G10、A5的单克隆抗体腹水做1:100稀释与LDH、CK、CKMB和GOT等心肌酶均无交叉反应。5株McAb的腹水效价为3.2×10-6~1.6×10-7。McAb相加试验表明,A5和G3可识别不同的抗原表位。  相似文献   

3.
应用常规方法建立了3株稳定分泌抗金黄色葡萄球菌C1型肠毒素(SEC1)单克隆抗体(McAb)的小鼠杂交瘤细胞系B3、C4和G8。其中B3和C4均为IgG1(k),G8为IgG2a(k)。B3和G8与SEA、SEB及SED均无交叉反应;C4虽与SEA和SED无交叉反应,但与SEB有交叉反应。间接ELISA测定小鼠腹水效价为10-5~10-8。应用识别不同表位的McAb建立了双McAb夹心ELISA法检测SEC1,敏感性可达1ng/ml。  相似文献   

4.
应用常规方法建立了4株稳定分泌抗重组人IL-6(rHuIL-6)单克隆抗体(McAb)的小鼠杂交瘤细胞系1H3、2A10、3A3和4B1。其中,1H3为IgG2b(k),2A10为IgG1(k),3A3和4B1为IgG2a(k)。4株McAb特异性强,与细胞因子IL-1β、IL-3、IL-8、TNF-α、GM-SCF、ICAM-1,以及受体菌菌体蛋白成分均无交叉反应。间接ELISA测定小鼠腹水McAb效价为10(-6)~10(-8)。应用识别不同表位的McAb建立了双抗体夹心ELISA法检测IL-6,敏感性为100pg/ml。初步应用表明可用于临床标本的检测。  相似文献   

5.
作者采用PharmaciaSephacrylS—300凝胶色谱柱,建立了IgG类McAb的一步法制各级纯化方法。该法是将McAb腹水直接上样,用pH7.410mmol/LPBS洗脱(流速0.5ml/min),即得到纯化的McAb。一次上样量40~50ml腹水,回收率为85%-90%.整个纯化周期4h。纯化的McAb经SDS—PAGE测定,纯度>90%,免疫组化ABC法测定活性为1:80000(7.8×10-11mol/L)。该法操作简单、快速,只要有一台核酸/蛋白检测仪,便可进行制备级水平的纯化。  相似文献   

6.
挑选了10株具有代表性的菌株,提取了它们的外膜蛋白(OMPs),SDS-PAGE分析发现:10株菌的OMPs带谱不完全相同,但都含有1,2/3,4和5类外膜蛋白,而且纯化的1类外膜蛋白(OMP1)显示一条分子量为(41或43)×10^3的蛋白带。从中选两株菌542852(B:NT:P1.2:L3,7,9:克隆I:RFLP-b20)和3407(B:15:P1.2:L3,7,9,克隆群I:RFLP-b  相似文献   

7.
凝胶色谱法半制备级纯化单克隆抗体   总被引:1,自引:0,他引:1  
作者采用SePllacylS-200凝胶色谱柱,在快速蛋白液相色谱系统(FPLC)建立了肝癌相关抗原单克隆抗体半制各级纯化方法。该法是将McAb腹水经40%饱和硫酸铵盐析粗分离后,再用凝胶色谱纯化,一次上样量相当于5~10ml腹水,可得纯化McAb30~60mg,得率为6~6.5mg/ml腹水,回收率为63%,一次色谱纯化周期2h,整个分离纬化可在3h内完成。经SDS—PAGE和DEAE离子交换色谱法检测McAb纯度大于80%,ABC染色法测定McAb活性为,1:40000(1.56×10-10mol/L),是一般实验室纯化IgG类McAb的实用方法。  相似文献   

8.
高效凝胶色谱一步法制备级纯化单克隆抗体   总被引:2,自引:0,他引:2  
作者采用Pharmacia Sephacryl S-300凝胶色谱柱,建立了IgG类McAb的一步法制备级纯化方法。该法是将McAb腹水直接上样,用pH7.410mmol/L PBS洗脱(流速0.5ml/min),即得到纯化的McAb。一次上样量40 ̄50ml腹水,回收率为85% ̄90%。整个纯化周期4h。纯化的McAb经SDS-PAGE测定,纯度〉90%,免疫组化ABC法测定活性为1:80000  相似文献   

9.
金葡菌C1型肠毒素免疫BALB/c小鼠,取其脾细胞与Sp2/0小鼠骨髓瘤细胞融合选出了能分泌高滴度McAb的18个克隆株,对其中10株进行了鉴定,6株属IgG1,4株属IgG3。用双抗体夹心法和特异性中和抑制试验比较了McAb和PcAb的敏感性和特异性,并用制备的McAb诊断试剂对SECI污染食品进行了检测应用,可特异检出lugSEC1/0.5g食物/ml。  相似文献   

10.
作者采用DEAE-40HR阴离子交换柱。在快速蛋白液相色谱系统(FPLC)建立了IgG类单克隆抗体制备级纯化方法。该法是将McAb腹水经50%饱和硫酸铵盐析粗分离后,再用阴离子交换色谱纯化,一次上样量相当于80~100ml腹水。可得纯化McAb500~600mg,得率为6~7mg/ml腹水,回收率为57~67%.一次纯化周期仅需45min。经SDS-PAGE检测McAb纯度为95%,ABC染色法测定McAb活性为1:20000(3.13×10-10mol/L).  相似文献   

11.
Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc2 (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.  相似文献   

12.
Renal dysplasia and asplenia in two sibs   总被引:2,自引:0,他引:2  
A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.  相似文献   

13.
About 1900, modern food selection and processing caused widespread epidemics of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a panhypovitaminosis B, i.e., a mixture of substrate beriberi and substrate pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse endemic disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for primates. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.  相似文献   

14.
15.
Newton H 《Medical history》2011,55(2):153-182
Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.  相似文献   

16.
Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.  相似文献   

17.
HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.  相似文献   

18.
The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.  相似文献   

19.
Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.  相似文献   

20.
Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号