4.5 Article

Identification of a novel single chain fragment variable antibody targeting CD24-expressing cancer cells

期刊

IMMUNOLOGY LETTERS
卷 190, 期 -, 页码 240-246

出版社

ELSEVIER SCIENCE BV
DOI: 10.1016/j.imlet.2017.08.028

关键词

-

资金

  1. Shahid Beheshti University of Medical Sciences [4967]

向作者/读者索取更多资源

CD24 is a heavy glycosylated protein that is overexpressed in numerous cancer and cancer stem cells. It is involved in the development, invasion, and metastasis of the cancer cells. CD24 can be considered in targeted cancer therapy as a new target. Here, phage display technology was used for the identification of novel scFv antibodies against CD24. To do so, the CD24 protein was expressed and purified from a stable transgenic CHO cell line. The cells were developed by using a CD24 encoding construct, which targets the 18S rRNA gene. The recombinant CD24 was used in the biopanning process. Four rounds of biopanning were performed with the Tomlinson J library. The polyclonal phage ELISA and DNA sequencing of the selected colonies confirmed that specific binders have been enriched during the biopanning process. Based on the DNA sequencing results, three phage-displaying scFv were obtained and their specificity to CD24 was verified in the monoclonal phage ELISA. The results showed that clone 3 (called J(d3)) has the highest affinity to CD24 and also is the most abundant clone (80%) among the three clones. Therefore, J(d3) was selected for further analyses. In order to produce soluble J(d3) scFv, the phage was transfected to E. coli BL21 pLysS. Soluble J(d3) scFv was expressed and purified successfully. The soluble J(d3) scFv showed appropriate affinity to recombinant CD24 in the ELISA analysis. Also, the immunocytochemistry experiment showed that the purified J(d3) scFv can bind to the CD24 expressing A549 cells. So, the scFv may be useful in the targeted delivery of drugs or diagnostic nanoparticles to the CD24 expressing cancer cells. To the best of our knowledge, this study is the first report to identify an scFv antibody against CD24, using the phage display method.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.5
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据