De novo protein sequencing, humanization and in vitro effects of an antihuman CD34 mouse monoclonal antibody

抗人 CD34 鼠单克隆抗体的从头蛋白质测序、人源化及体外作用

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作者:Chia-Yu Fan, Sheng-Yu Huang, Min-Yuan Chou, Ping-Chiang Lyu

Abstract

QBEND/10 is a mouse immunoglobulin lambda-chain monoclonal antibody with strict specificity against human hematopoietic progenitor cell antigen CD34. Our in vitro study showed that QBEND/10 impairs the tube formation of human umbilical vein endothelial cells (HUVECs), suggesting that the antibody may be of potential benefit in blocking tumor angiogenesis. We provided a de novo protein sequencing method through tandem mass spectrometry to identify the amino acid sequences in the variable heavy and light chains of QBEND/10. To reduce immunogenicity for clinical applications, QBEND/10 was further humanized using the resurfacing approach. We demonstrate that the de novo sequenced and humanized QBEND/10 retains the biological functions of the parental mouse counterpart, including the binding kinetics to CD34 and blockage of the tube formation of the HUVECs.

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