Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5507074 | Biochemistry and Biophysics Reports | 2017 | 10 Pages |
â¢A method of de novo protein sequencing of mouse mAb was developed.â¢Humanization of a mouse monoclonal antibody has been performed.â¢Antihuman CD34 antibody impairs the tube formation of HUVECs.â¢CD34 could be a potential drug target for antiangiogenic therapy.
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.