Page 138 - ebook

[B. Cell Biology/Stem Cell] B-4

Evaluation of novel anti-human CD133 human antibody

developed using phage display in human synthetic scFv

library

Sang-In Park¹, Do-Hyung Kim¹, Sungjin Lee¹, Hea Li Ko¹, Deuk-ki Lee¹, Seok-Ho Go¹, Younghyeon Kim²,

Jin Won Park¹*

¹Division of research program, Scripps Korea Antibody Institute, Chuncheon-si 24341, Korea, ²Department of

Systems Immunology, College of Biomedical science, Kangwon National University, Chuncheon-si 24341, Korea

CD133 is colon cancer stem cell surface marker and highly expressed in refractory colorectal cancers. We developed
novel anti-human CD133 antibody using phage display technics in human synthetic scFv library. In this study, novel

CD133 human antibody evaluated recombinant CD133 antigen and live cell surface binding affinity. Antibody
screening were performed 3 times by phage display using as antigen recombinant human CD133 produced from

293 cell. A scFv was converted to IgG form through gene cloning. Antibody binding affinities to CD133 were
analyzed by ELISA, flow cytometry. CD133 antibody screenings were performed three times with phage display, and

the final titers were 1.08x108 CFU/ml. Each 400 colonies picked up and confirming by ELISA. We selected 15 effective
scFv antibodies with individual sequences, among them, picked 6 lead scFv antibodies with flow cytometry analysis.

After human IgG form conversions, three CD133 antibodies were showed high affinities to HT29 cell line high CD133
expressed cell line on flow cytometry analysis. We developed three novel anti-human CD133 human antibody that

showed high affinity on high CD133 expressed cell line.