Combine Phage Antibody Display Library Selection on Patient Tissue Specimens with Laser Capture Microdissection to Identify Novel Human Antibodies Targeting Clinically Relevant Tumor Antigens.-猫眼课题宝

将患者组织样本上的噬菌体抗体展示库选择与激光捕获显微切割相结合，以识别针对临床相关肿瘤抗原的新型人类抗体。

DOI：

10.1007/978-1-4939-7447-4_18

发表时间：

2018

期刊：

Methods in molecular biology (Clifton, N.J.)

影响因子：

通讯作者：

中科院分区：

文献类型：

作者：

Su,Yang;Bidlingmaier,Scott;Lee,Nam-Kyung;Liu,Bin

A functional approach to generate tumor-targeting human monoclonal antibodies is through selection of phage antibody display libraries directly on tumor cells. Although technically convenient, the use of cancer cell lines for the selection has limitations as those cell lines often undergo genetic and epigenetic changes during prolonged in vitro culture and alter their cell surface antigen expression profile. The key is to develop a technology that allows selection of phage antibody display libraries on tumor cells in situ residing in their natural tissue microenvironment. Laser capture microdissection (LCM) permits the precise procurement of tumor cells from human cancer patient tissue sections. Here, we describe a LCM-based method for selecting phage antibodies against tumor cells in situ using both fresh frozen and paraffin-embedded tissues. To restrict the selection to antibodies that bind internalizing epitopes, the method utilizes a polyclonal phage population pre-enriched for internalizing phage antibodies. The ability to recognize tumor cells in situ residing in their natural tissue microenvironment and to deliver payload intracellularly makes these LCM-selected antibodies attractive candidates for the development of targeted cancer therapeutics.