单纯疱疹病毒荧光标记单克隆抗体的制备及临床应用  

Preparation and Preliminary Application of Fluorescein Labelled Monoclonal Antibodies against Herpes Simplex Virus

作　　者：肖庆忠[1] 陈柏铭[1] 宁波[2] 

机构地区：[1]广州医学院第一附属医院检验科,广州510120 [2]中山医科大学免疫学教研室

出　　处：《广州医学院学报》2002年第1期8-12,共5页Academic Journal of Guangzhou Medical College

摘　　要：目的:为了了解所制备的单纯疱疹病毒荧光标记单克隆抗体的临床应用价值。方法:本研究利用杂交瘤技术制备了多株抗单纯疱疹病毒型共有及型特异性单克隆抗体(６Ｂ３－２－５、Ｄａ５,５Ｂ４－６),并通过ＢＡＬＢ/Ｃ鼠体内诱生制备了较大量的单抗,经去脂澄清处理,利用辛酸－硫酸铵二步法纯化了单抗,经ＳＤＳ－ＰＡＧＥ及薄层扫描分析显示其纯度;在ｐＨ９．０－９．５条件下,采用低温袋内透析法标记了单抗５Ｂ４－６及６Ｂ３－２－５,然后经透析及ＳｅｐｈｅｄｅｘＧ－５０柱去除游离荧光素;标记单抗采用直接免疫荧光法(ＤＦＡ)检测各类临床标本１１０份及正常标本２６份,并与间接免疫荧光法(ＩＦＡ)及组织培养分离法作了初步比较。结果:１．纯化后的各单抗在解链状态下为二条链,一条为轻链,分子量各为 ２０．６ ＫＤ(５Ｂ４－６)、２４．６ ＫＤ(６Ｄ３－２－５)及 ２６．１ＫＤ(Ｄａ５),一条为重链,分子量各为５７．８ ＫＤ(５Ｂ４－６)、５４．８ ＫＤ(６Ｂ３－２－５)及５２．７ＫＤ(Ｄａ５),由此推算出其单抗分子量各为１５７ ＫＤ、１５９ ＫＤ、１５８ ＫＤ。纯化回收率各为４３．７７%、４７．２１%及 ３８．８%。２．标记单抗 Ｆ/Ｐ值各为ｌ．４ 及１．４６,利用(ＤＦＡ)法测定其效价各为 １:３２?Activity measurements of serum 5'-nucleotidase were carried out in 138 cases suffering from different kinds of liver diseases. Results showed elevations of the enzyme activities in varying degrees among different liver disorders, In liver cancer, a 100％ positive rate was observed, but there was no elevation of the enzyme activity in extrshepatic cancers. A preliminary impression obtained from our studies is that serum 5' nucleotidase activity may be used as a reference index for differential diagnosis between hepatic and extrahepatic cancers.

关 键 词：单纯疱疹病毒 单克隆抗体 微量免疫荧光法 荧光标记 

分 类 号：R446.61[医药卫生—诊断学]