De novo protein sequencing, humanization and in vitro effects of an antihuman CD34 mouse monoclonal antibody
peer-reviewed · Biochemistry and Biophysics Reports · 2017
| Date | 2017-03-01 |
| Type | peer-reviewed |
| Venue | Biochemistry and Biophysics Reports |
| Publisher | Elsevier BV |
| Contribution | downstream-application |
| DOI | 10.1016/j.bbrep.2016.11.006 |
| Citations (OpenAlex) | 2 |
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.
Methods and tools
- Anti-CD34 mAb sequencing and humanization: De novo protein sequencing of QBEND/10, a mouse monoclonal against the haematopoietic progenitor antigen CD34, followed by humanization and in vitro testing of the resulting antibody on endothelial tube formation.