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1.
以基因工程人表皮生长因子(rhEGF)为抗原,免疫BALB/c小鼠,将免疫的小鼠脾细胞与小鼠Sp2/0-Ag14骨髓瘤细胞进行融合,经多次筛选和再克隆化,获得3株能稳定分泌抗rhEGF单克隆抗体(McAbs)的杂交瘤细胞株.对该3株杂交瘤细胞小鼠腹水进行了抗体效价测定,结果显示在1×10-6~2×10-7之间.对上述3种杂交瘤单抗(分别命名为:AE-2A4,AE-2G5和AE-3E11)进行了特异性、抗原决定簇及相对亲和力测定.结果表明,它们都具有抗rhEGF高特异性;并分别抗两个不同抗原决定簇;相对亲和力大小次序为:AE-3E11>AE-2A4>AE-2G5.Ig亚类鉴定结果显示:AE-2A4,AE-3E11同为IgG1亚类,而AE-2G5为IgG2b亚类.确定AE-2A4单克隆抗体可用于制备高纯rhEGF.  相似文献   

2.
利用细胞融合技术建立了5株分泌抗温和气单胞菌CR79-1-1株单克隆抗体(McAb)的杂交瘤细胞系,通过特异性鉴定获得两株与参考菌株中的温和气单胞菌发生反应,但与嗜水气单胞菌以及鳗弧菌、爱德华氏菌、克鲁氏耶尔森菌、大肠杆菌、荧光假单胞菌均不反应的杂交瘤细胞系,分别命名为2G3和1A4.快速酶联免疫分析法(ELISA)测得2G3和1A4的亲和常数分别为1.72×108和5×108M-1;应用方阵配对实验证实,2株单抗针对特异性抗原上不同表位.  相似文献   

3.
用灭活迟缓爱德华氏菌菌株AL60306NA1免疫Balb/c小鼠,采用杂交瘤技术制备抗迟缓爱德华氏菌的McAb,获得3株可持续分泌抗迟缓爱德华氏菌McAb的杂交瘤细胞株3A7、4F11和4C9;腹水效价分别达到1.0×10-6、1.0×10-6和1.0×10-5;细胞亚型分别为IgG1、IgG2b、IgM.交叉反应结果...  相似文献   

4.
褪黑素单克隆抗体的制备及鉴定   总被引:1,自引:0,他引:1  
为了获得抗褪黑素(MT)高效价的单克隆抗体,用甲醛将MT连结到BSA上,然后用连结物背部皮下多点免疫Balb/c小鼠,用PEG诱导免疫鼠脾细胞与Sp2/0细胞融合,经ELISA法筛选阳性克隆株及测定效价,用抗体与同类物之间的交叉反应进行特异性鉴定.最后获得5株(4C9D7、3E12C7H11E7、3E12A9D7、6A6A3A2C3、1E1E2H2H6)能稳定分泌高效价抗MT的单克隆抗体(McAb)的杂交瘤细胞。  相似文献   

5.
心肌肌钙蛋白I-C复合物酶联免疫检测方法的建立   总被引:2,自引:0,他引:2  
为了制备抗人心肌肌钙蛋白I(cTnI)单克隆抗体、建立检测人cTnI链酶亲和素-生物素双抗体夹心酶联免疫吸附测定(ELISA)方法,用基因工程表达的人cTnI免疫雌性BALB/c小鼠,以杂交瘤技术制备得到三株特异性抗人cTnI的单克隆抗体记为6G3,6A6,3G9.三株杂交瘤细胞中6G3和6A6为IgG1,3G9为IgG2b,腹水效价分别为4×10-5,4×10-5和10-5.亲和常数为5.0×109,6.4×109和3.8×109mol-1.Westernblotting结果表明6G3和6A6能够识别cTnI-C复合物.把6G3抗体生物素化并制备抗cTnI、cTnC和cTnI-C的大鼠多克隆抗体.以多抗作为固相捕捉抗体,生物素化6G3抗体作为检测抗体,建立检测cTnI的双抗体夹心ELISA方法并初步应用于临床病人标本检测.以cTnI-C复合物多抗捕捉检测方法具有良好的灵敏度、特异性和准确性,灵敏度为0.9ng/mL,较包被其他抗体捕捉方法提高1~2倍.批内变异系数为5.03%,回收率为94.56%.测定17例AMI病人和18例血清样本临床诊断结果符合率为100%.  相似文献   

6.
本文运用B淋巴细胞杂交瘤技术得到了4株稳定分泌抗hGH羊克隆抗体的杂交瘤细胞株,并对其特性进行了鉴定。其腹水滴度高达0.6~10×10~5,亲和常数在0.53~9×10~9 M~(-1)之间。4种单克隆抗体的相应抗原决定簇分属于 hGH 分子表面3个不同区域;除1种单克隆抗体与 hPL 有交叉反应外,其余与hPL和hPRL皆无交叉反应。另外,本文采用的微量免疫法也有一定的实用价值。  相似文献   

7.
以合成肽COX-2-KLH为完全抗原免疫BALB/c 小鼠,通过脾细胞与骨髓瘤细胞SP2/0融合的杂交瘤技术,筛选获得4株(1E10、3D6、5B11、6G4)稳定分泌抗COX-2单克隆抗体(McAb)的杂交瘤细胞株.采用ELISA的方法鉴定McAb效价、亚型及特异性.各株McAb均属IgG1 亚型,其中1E10腹水纯化后MCAb效价在1∶107以上,SDS-PAGE电泳为单一条带,与同源异构体蛋白不发生交叉反应.获得的McAb被成功应用于COX-2生物学研究中的免疫组织化学(IHC)和Western Blot试验中.本研究获得的单抗具有特异、高效的特点,为COX-2生物学相关研究提供理论依据.  相似文献   

8.
为了对伊氏锥虫病的免疫诊断提供有价值的单克隆抗体,笔者通过了3次SP2/0骨髓瘤细胞与伊氏锥虫抗原免疫的BALB/c小鼠脾细胞间的融合试验和多次筛选以及克隆化,建立起7株分泌单克隆抗体的杂交瘤细胞株。对其中ⅡG_6杂交瘤作了进一步的检定分析,用ⅡG_6上清液对T.e.抗原进行ELISA试验,显示出高度的特异性,与血吸虫、弓形虫抗原均未出现交叉反应。ELISA测试ⅡG_6细胞株培养上清效价为1:5.1×10~3,而杂交瘤细胞接种BALB/c小鼠诱生腹水抗体的效价为1:1.6×10~7。该株单克隆抗体免疫球蛋白的类型鉴定属IgG_1亚类。该株细胞已在体外培养下,稳定地分泌特异性抗体至少达6个月之久,同时在液氮冻存近8个月之后仍能保持分泌抗体的能力。  相似文献   

9.
目的 研究甘油-3-磷酸对甲肝疫苗诱导的小鼠体液免疫应答的影响。方法 选取49只雌性ICR小鼠,分7组,每组7只,这7组小鼠分别为阴性对照、单纯抗原组、铝佐剂组和4组甘油-3-磷酸组,分别在免疫之后的第4、8、12、16、20周用酶联免疫法(ELISA)测定小鼠血清中的特异性的anti-HAV IgG水平。结果 在小鼠免疫之后的第4、8、12、16、20周单纯抗原组、铝佐剂组和4组不同剂量甘油-3-磷酸组均能检测到特异性抗HAV抗体,并且趋势为先升高后降低,且在免疫后的第8周最高;第4周和第8周的甘油-3-磷酸实验组的结果均高于单纯抗原组,且差异均具有统计学意义(P<0.05),并且第4周甘油-3-磷酸组抗体水平均超过铝佐剂组,并具有统计学意义(P<0.05),其中甘油-3-磷酸2 mg为最佳剂量组,其抗体水平在免疫后的第4、8、12、16、20周均高于单纯抗原组,且在第8周抗体水平达到峰值,抗HAV IgG水平为lg(3.064±0.07851),铝佐剂的也是在第8周抗体水平达到峰值,抗HAV IgG水平为lg(3.150 ±0.1382)。结论 甘油-3-磷酸有免疫佐剂的作用,能够有效、迅速地增强HAV诱导的体液免疫应答,是一种潜在、安全有效的新型疫苗佐剂。  相似文献   

10.
极谱法连续测定痕量铅和镉   总被引:1,自引:0,他引:1  
研究了Pb2+和Cd2+与KCI-酒石酸钠-三乙醇胺-明胶体系的二次导数极谱波.在pH=4.5的HAc-NaAc介质中,Pb2+和Cd2+分别于-0.46V和-0.64V电位处产生一良好的极谱波.峰电流与Pb2+和Cd2+的浓度分别在1×10-5~3×10-1 g·L-1和5×10-5~6×10-3 g·L-1范围内呈线性关系.Pb2+和Cd2+的检出限分别为1×10-7 g·L-1和5×10-7 g·L-1.本法准确、简便、快速、选择性高.已直接用于饮料中铅和镉的连续测定,同时也对酒及面粉样品进行了测定,回收率分别为99.9%~100.1%和97.0%~104.8%.  相似文献   

11.
Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.  相似文献   

12.
何延凌 《科技信息》2008,(4):258-258
Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women's language from these perspectives: pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women's language, more attention should be paid to language use in social context. What's more, the linguistic phenomena in a speaking community can be understood more thoroughly.  相似文献   

13.
理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。  相似文献   

14.
As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.  相似文献   

15.
正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-  相似文献   

16.
We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-  相似文献   

17.
正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that  相似文献   

18.
For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the  相似文献   

19.
<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with  相似文献   

20.
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