De novo protein sequencing,humanization and in vitro effects of an antihuman CD34 mouse monoclonal antibody |
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Authors: | Chia-Yu Fan Sheng-Yu Huang Min-Yuan Chou Ping-Chiang Lyu |
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Affiliation: | 1. Institute of Bioinformatics and Structural Biology & Department of Life Science, National Tsing Hua University, Hsinchu, Taiwan;2. Biomedical Technology and Device Research Laboratories, Industrial Technology Research Institute, Hsinchu, Taiwan;3. Mithra Biotechnology Inc., New Taipei City, Taiwan |
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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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Keywords: | Monoclonal antibody De novo sequencing Humanization CD34 Antiangiogenic therapy |
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