[1]吴梦雪,雷成明,谭祖稳,等.131I-RP215 McAb分子探针的制备、鉴定及荷人肺腺癌裸鼠放射免疫显像[J].第三军医大学学报,2018,40(07):584-589.
 WU Mengxue,LEI Chengming,TAN Zuwen,et al.Preparation and identification of 131I-RP215 McAb molecular probe and its application for radioimmunoimaging in lung adenocarcinoma-bearing nude mice[J].J Third Mil Med Univ,2018,40(07):584-589.
点击复制

131I-RP215 McAb分子探针的制备、鉴定及荷人肺腺癌裸鼠放射免疫显像(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第07期
页码:
584-589
栏目:
基础医学
出版日期:
2018-04-15

文章信息/Info

Title:
Preparation and identification of 131I-RP215 McAb molecular probe and its application for radioimmunoimaging in lung adenocarcinoma-bearing nude mice
作者:
吴梦雪雷成明谭祖稳张辉唐嘉励段东
重庆医科大学附属第一医院核医学科
 
Author(s):
WU Mengxue LEI Chengming TAN Zuwen ZHANG Hui TANG Jiali DUAN

DongDepartment of Nuclear Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
核素分子探针RP215单克隆抗体CA215肺腺癌放射免疫显像
Keywords:
nuclide molecule probe RP215 monoclonal antibodies CA215 lung adenocarcinoma radioimmunoimaging
分类号:
R392-33; R734.2; R817.4
文献标志码:
A
摘要:

目的    制备靶向肺腺癌细胞表面碳水化合物相关抗原(carbohydrate antigen 215,CA215)的核素分子探针131I-RP215单克隆抗体(monoclonal antibody,McAb),鉴定其理化性质并探讨其在荷瘤裸鼠模型中的体内分布情况和放射免疫显像特点。 方法    采用氯胺T法对McAb RP215进行131I标记,Sephadex G25M 柱纯化标记抗体,并鉴定其理化特性,观察该抗体标记物在荷A549裸鼠模型中的生物分布及放射免疫显像情况。 结果    131I-RP215 McAb 标记率为(91.03±2.36)%,纯化后的放化纯度为(93.15±1.40)%,比活度(3.37±0.42)MBq/μg;具有较好的稳定性及免疫活性。荷瘤裸鼠体内生物分布及SPECT显像结果显示,131I-RP215 McAb能够在肿瘤组织中选择性浓聚,131IRP215 McAb作用24 h时,肿瘤/肌肉比达最高,且瘤体显影最清晰。结论    成功制备了特性理想的 131I-RP215 McAb分子探针,在荷人肺腺癌裸鼠模型中有较好的显像效果。

Abstract:

Objective     To prepare a targeted probe for nuclide labeling of 131I-RP215 monoclonal antibody (McAb) against carbohydrate-associated antigen, carbohydrate antigen 215 (CA215) which are expressed on the surface of lung adenocarcinoma cells, identify the physico-chemical properties of the probe, and to explore its distribution in vivo and radioimmunoimaging (RII) characteristics in tumor-bearing nude mice model. Methods    McAb RP215 was labeled with 131I by the chloramine T method, and then the products were purified with the Sephadex G25M column. The physico-chemical properties of the obtained McAbs were studied. Nude mice model bearing human lung adenocarcinoma was established by transplantation of A549 cells. Then the mice received the injection of 131I-RP215 McAbs via tail vein, and the bio-distribution and RII characteristics were observed with single-photon emission computerized tomography (SPECT). Results     The obtained 131I-RP215 McAbs showed a labeling rate of (91.03±2.36)%, a radiochemical purity of (93.15±1.40)%, and a specific activity of 3.37±0.42 MBq/μg, and had good stabilities and immunological competence. Biodistribution and SPECT imaging showed 131I-RP215 selectively accumulated in tumor tissue, and the target/non-target (T/NT) ratio reached the highest with the clearest tumor imaging at 24 h after injection of McAbs. Conclusion    Molecular probes, 131I-RP215 McAbs, with ideal physicochemical properties are successfully prepared, and show satisfactory SPECT imaging in tumor-bearing nude mice.

参考文献/References:

[1]LEE G, ZHU M, GE B, et al. Widespread expressions of immunoglobulin superfamily proteins in cancer cells[J]. Cancer Immunol Immunother, 2012,61(1): 89-99. DOI: 10.1007/s00262-011-1088-1.
[2]LEE G. Cancer cellexpressed immunoglobulins: CA215 as a pan cancer marker and its diagnostic applications[J]. Cancer Biomark, 2009,5(3):137-142. DOI: 10.3233/CBM-2009-0610.
[3]LEE G, LAFLAMME E, CHIEN C H, et al. Molecular identity of a pan cancer marker, CA215[J]. Cancer Biol Ther, 2008,7(12): 2007-2014. DOI: 10.4161/cbt.7.12.6984.
[4]LEE G,WU Q,LI C H, et al. Recent studies of a new carbohydrate-associated pan cancer marker, CA215[J]. J Clin Ligand Assay, 2006,29(1): 47-51.
[5]LEE G, GE B, HUANG T K, et al. Positive identification of CA215 pan cancer biomarker from serum specimens of cancer patients[J]. Cancer Biomark, 2010, 6(2): 111-117. DOI:10.3233/CBM-2009-0134.
[6]LEE G, AZADI P. Peptide mapping and glycoanalysis of cancer cellexpressed glycoproteins CA215 recognized by RP215 monoclonal antibody[J]. J Carbohyd Chem, 2012,31(1): 10-30.
[7]LIAO Q, LIU W, LIU Y, et al. Aberrant high expression of immunoglobulin G in epithelial stem/progenitor-like cells contributes to tumor initiation and metastasis[J]. Oncotarget, 2015, 6(37):40081-40094. DOI: 10.18632/oncotarget.5542.
[8]LEE G, CHEUNG A P, LI B, et al. Molecular and immuno-characteristics of immunoglobulin-like glycoproteins in cancer cellexpressed biomarker,CA215[J]. Immunol Invest,2012,41(4):429-446. DOI:10.3109/08820139.2012.661007.
[9]LEE G, ZHU M, GE B. Potential monoclonal antibody therapy for the treatment of ovarian cancer[M]//FARGHALY S A.Ovarian cancer—basic science perspective. Rijeka:Tech-Open Access Publisher,  2012:385-406.
[10]LEE G, GE B. Inhibition of in vitro tumor cell growth by RP215 monoclonal antibody and antibodies raised against its antiidiotype antibodies[J]. Cancer Immunol Immunother,2010,59(9):1347-1356.DOI:10.1007/s0026201008647.
[11]LEE G, CHU R A, TING H H. Preclinical assessment of anti-cancer drugs by using RP215 monoclonal antibody[J]. Cancer Biol Ther, 2009, 8(2):161-166. DOI:10.4161/cbt.8.2.7117.
[12]LIU Y, LIU D, WANG C, et al. Binding of the monoclonal antibody RP215 to immunoglobulin G in metastatic lung adenocarcinomas is correlated with poor prognosis[J]. Histopathology, 2015, 67(5):645-653. DOI:10.1111/his.12686.
[13]LEE G, GE B. Cancer cell expressions of immunoglobulin heavy chains with unique carbohydrateassociated biomarker[J]. Cancer Biomark, 2009, 5(4): 177-188. DOI:10.3233/CBM20090102.
[14]HUDSON P J, SOURIAU C. Engineered antibodies[J]. Nat Med, 2003, 9(1):  129-134. DOI:10.1038/nm0103-129.
[15]REITER Y, SCHUCK P, BOYD L F, et al.An antibody single-domain phage display library of a native heavy chain variable region: isolation of functional single-domain VH molecules with a unique interface[J]. J Mol Biol, 1999, 290(3):685-698. DOI:10.1006/jmbi.1999.2923.
[16]YU F, WANG Y, XIAO Y, et al. RP215 single chain fragment variable and single domain recombinant antibodies induce cell cycle arrest at G0/G1 phase in breast cancer[J].Mol immunol, 2014,59(1):100-109. DOI:10.1016/j.molimm.2014.01.007.
[17]庞华. 肺腺癌人源噬菌体单链抗体制备及其在荷人肺腺癌裸鼠模型中的放免显像[D]. 重庆:重庆医科大学, 2008.
PANG H. The construction of human single-chain variable fragment antibodies of lung adenocarcinoma and its radioimmunoimaging in bearing cancer nude mice[D]. Chongqing: Chongqing Medical University,2008.
[18]王洁, 李建国, 庞华,等. 抗HIF-1α肺腺癌人源单链抗体的制备和放射免疫显像[J]. 第三军医大学学报,2010,32(17):1851-1853.DOI: 10.16016/j.10005404. 2010.17.033.
WANG J, LI J G, PANG H, et al. Preparation of human anti-HIF-1α scFv of lung adenocarcinoma and its radioimmunoimaging[J]. J Third Mil Med Univ, 2010, 32(17): 1851-1853. DOI: 10.16016/j.10005404.2010. 17.033.  

更新日期/Last Update: 2018-04-10