Identification and application of fluorescent labeled small molecule antibodies based on cell surface specific recognition

ZHANG Jing; XIONG Ming; LI Xiaojuan; MA Wei; ZHANG Dajin; QIAO Yuanyuan

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Medical Journal of the Chinese People Armed Police Forces ›› 2021, Vol. 32 ›› Issue (4) : 304-308.

Identification and application of fluorescent labeled small molecule antibodies based on cell surface specific recognition

ZHANG Jing¹, XIONG Ming², LI Xiaojuan¹, MA Wei², ZHANG Dajin², QIAO Yuanyuan²

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Abstract

Objective To establish a method of identification of fluorescent-labeled small molecule antibodies based on cell surface specific recognition and to verify their applicability in detection of peripheral blood circulating tumor cells (CTCs) of esophageal cancer.Methods Phage antibodies NFC 20b and NFC 70a were obtained via high-throughput screening and enrichment using phage display technology in the early stage of this study. After fluorescence labeling, the aggregation tracer between them and esophageal cancer cells was observed in zebrafish. Small molecule antibodies were also used to detect CTCs in peripheral blood of esophageal cancer patients. Meanwhile, the expression profile of antigen proteins was analyzed using the high-throughput protein chip.Results The distribution of fluorescent labeled small molecule antibodies NFC 20b and NFC 70a could be observed in the dorsal aorta, caudal artery and caudal vein of zebrafish, so was the binding of antibodies to transplanted tumors in yolk sac. The protein chip used to screen the expression profile of the antigen protein proved that the antigen protein bound by NFC 20b was cyclin dependent kinase 2(CDK2).Conclusions Small molecule antibodies labeled by fluorescence labeling technology can be distributed to yolk sac through blood circulation, and can aggregate with and bind to transplanted tumors of esophageal cancer in yolk sac. This approach can be used to detect CTCs in peripheral blood of ESCC. The protein chip can be used to assist in identifying the antigen binding to small antibodies.

Key words

circulating tumor cells / zebrafish / fluorescent labeled / antibody / protein chip

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ZHANG Jing, XIONG Ming, LI Xiaojuan, MA Wei, ZHANG Dajin, QIAO Yuanyuan. Identification and application of fluorescent labeled small molecule antibodies based on cell surface specific recognition[J]. Medical Journal of the Chinese People Armed Police Forces. 2021, 32(4): 304-308

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