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

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.

SUBMITTER: Fan CY

PROVIDER: S-EPMC5614552 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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

Fan Chia-Yu CY Huang Sheng-Yu SY Chou Min-Yuan MY Lyu Ping-Chiang PC

Biochemistry and biophysics reports 20161118

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 an ...[more]

PMID: 28955989

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