Prokaryotic expression of MLAA-34 and generation of a novel human ScFv against MLAA-34 by phage display technology
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Yang Zhang1, Pengyu Zhang1, Aili He1, Yun Yang1, Jianli Wang1, Hui Zhang1 and Wanggang Zhang1
1Department of Hematology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xi’an, Shaanxi 710004, P.R. China
Wanggang Zhang, email: [email protected]
Keywords: MLAA-34, acute monocytic leukemia, phage antibody library, single chain antibody fragment, antibody-based therapy
Received: December 06, 2016 Accepted: March 09, 2017 Published: March 28, 2017
MLAA-34 is a newly identified monocytic leukemia-associated antigen that is overexpressed in acute monocytic leukemia specifically, thus providing a novel target for the therapy of acute monocytic leukemia. In this study, we first expressed MLAA-34 protein in Escherichia coli (E.coli) BL21 (DE3) cells and purified it by nickel ion affinity chromatography with high purity (>90%). Then, MLAA-34 was used as antigen for biopanning anti-MLAA-34 single chain antibody fragment (ScFv) from a fully human ScFv library, and a high affinity ScFv named MA1 was selected by phage-ELISA. Finally, after expression of MA1, we found that MA1 can specifically bind with U937 MLAA-34 positive cells, and the binding affinity of MA1 was at the nanomolar level. Furthermore, inhibition of U937 cell proliferation indicated that the novel antibody MA1 has the potential to be used as a therapeutic agent for acute monocytic leukemia.
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