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[1]李先富,潘秀珍,赵华梅,等.2型猪链球菌谷氨酸脱氢酶单克隆抗体的制备与鉴定[J].医学研究与战创伤救治(原医学研究生学报),2010,12(05):385-387.
　LI Xian-fu,PAN Xiu-zhen,ZHAO Hua-mei,et al.Preparation and identification of monoclonal antibodies against GDH of Streptococcus suis serotype 2 highly virulent strain isolated from china[J].JOURNAL OF MEDICALRESEARCH —COMBAT TRAUMA CARE,2010,12(05):385-387.
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2型猪链球菌谷氨酸脱氢酶单克隆抗体的制备与鉴定()
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《医学研究与战创伤救治》（原医学研究生学报）[ISSN:1672-271X/CN:32-1713/R]

卷:
第12卷
期数:
2010年05期
页码:
385-387
栏目:
出版日期:
2010-09-20

文章信息/Info

Title:
Preparation and identification of monoclonal antibodies against GDH of Streptococcus suis serotype 2 highly virulent strain isolated from china
文章编号:
1672-271X(2010)05-0385-04
作者:
李先富; 潘秀珍; 赵华梅; 王长军; 唐家琪
210002 江苏南京，南京军区疾病预防控制中心流行病微生物学研究所
Author(s):
LI Xian-fu;  PAN Xiu-zhen;  ZHAO Hua-mei;  WANG Chang-jun;  TANG Jia-qi.
Centers for Disease Control and Prevention of Nanjing Military Command，Nanjing，Jiangsu 210002，China
关键词:
猪链球菌; 谷氨酸脱氢酶; 单克隆抗体
Keywords:
Streptococcus suis;  glutamate dehydrogenase;  monoclonal antibody
分类号:
R378.1+2
DOI:
-
文献标志码:
A
摘要:
目的 制备抗猪链球菌(Streptococcus suis，S. suis)谷氨酸脱氢酶(glutamate dehydrogenase, GDH)的单克隆抗体。方法 分别以2型S. suis全菌细胞和重组GDH蛋白为免疫原免疫BALB/c小鼠；采用淋巴细胞杂交瘤技术将免疫后的小鼠脾细胞与SP2/0骨髓瘤细胞融合；酶联免疫吸附试验(ELISA)筛选分泌抗GDH单克隆抗体的杂交瘤细胞株并克隆化；免疫印迹(Western blot)鉴定GDH单克隆抗体的特异性；制备小鼠腹水并进行抗体效价测定。结果 细胞融合后经ELISA筛选获得分泌的阳性细胞株，经过有限稀释克隆化筛选，获得4株能稳定分泌抗GDH单克隆抗体的杂交瘤细胞株。单抗腹水的抗体效价均在1∶102 400以上。结论 本研究获得了4株能稳定分泌抗体且其分泌的抗体能够与GDH蛋白进行特异性反应的单克隆抗体细胞株，所获得特异性单克隆抗体能用于猪链球菌结构和功能的研究，也能够进一步用于猪链球菌感染血清学诊断方法研究。
Abstract:
Objective To prepare monoclonal antibodies (McAbs) against glutamate dehydrogenase (GDH) protein of Streptococcus suis (S. suis).Methods S. suis isolates and the GDH recombinant protein were used as antigen to immunize female BALB/ c mice respectively.The immunized splenic cells of mice were fused with murine myeloma SP2/0 cells.The hybridoma cell strains secreting McAb against GDH were screened with enzyme linked immunosorbent assay (ELISA).The specificity of the secreted GDH McAbs was confirmed by Western blot.The ascites were harvested and the antibody titer was measured.Results Four hybridoma cell lines secreting McAb against GDH were obtained by fusion of mouse myeloma cells SP2/0 with splenic cells from BALB/c mice immunized with the purified expression product of GDH.Titers of McAbs in ascites were higher than 1∶102 400.Conclusion The McAbs against GDH were successfully prepared and identified, which would be useful for the detection of S. suis infection.

参考文献/References:

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[7]赵华梅，潘秀珍，王长军，等.猪链球菌2型谷氨酸脱氢酶基因的克隆表达及酶活性测定[J].中国人兽共患病学报,2006,22(1):22-25.
[8]潘秀珍,赵华梅,葛俊超, 等.信号肽与猪链球菌GDH 融合的DNA疫苗构建及体液免疫研究[J].免疫学杂志, 2008, 24(3): 311-314.
[9]潘秀珍，赵华梅，葛俊超，等.gdh基因在猪链球菌中的分布及重组GDH的抗原性研究[J].中国人兽共患病学报,2008,24(6):522-525.
[10]Okwumabua O, Persaud JS, Reddy PG.Cloning and characterization of the gene encoding the glutamate dehydrogenase of Streptococcus suis serotype 2[J].Clin Diagn Lab Immunol,2001,8(2):251-257.

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备注/Memo

备注/Memo:
国家自然科学基金项目(30730081，81071317)；江苏省自然科学基金项目(BK2007013)；南京军区医学科技创新课题(07Z045)；南京军区卫生专业人才培养“122工程”资助项目
更新日期/Last Update: 2010-09-20