title: Prokaryotic expression of MLAA-34 and generation of a novel human ScFv against MLAA-34 by phage display technology.
abstract: 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.
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Pengyu
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Aili
authors.initials: A
authors.lastname: He
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Jianli
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.
authors.forename: Wanggang
authors.initials: W
authors.lastname: Zhang
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology*; D051017:Apoptosis Regulatory Proteins / Q000276:immunology*; D049109:Cell Proliferation / Q000187:drug effects; D061505:Cell Surface Display Techniques / Q000379:methods*; D004926:Escherichia coli; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D007948:Leukemia, Monocytic, Acute; D057127:Single-Chain Antibodies; D020298:U937 Cells
chemical_list: D000951:Antigens, Neoplasm; D051017:Apoptosis Regulatory Proteins; C534238:CAB39L protein, human; D057127:Single-Chain Antibodies
keywords: MLAA-34; acute monocytic leukemia; antibody-based therapy; phage antibody library; single chain antibody fragment