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An antibody CDR3-erythropoietin fusion protein.

ABSTRACT: X-ray crystallographic analysis of a bovine antibody (BLV1H12) revealed a unique scaffold in its ultralong heavy chain complementarity determining region 3 (CDR3H) that folds into a solvent exposed, antiparallel ?-stranded "stalk" fused with a disulfide cross-linked "knob" domain. This unusual variable region motif provides a novel approach for generating chimeric antibodies with novel activities. Toward this end, human erythropoietin (hEPO) was substituted for the "knob" domain in this antibody to afford an antibody-hEPO (Ab-hEPO) fusion protein that efficiently expresses in mammalian cells. Ab-hEPO proliferated TF-1 cells with a potency comparable to that of hEPO (EC50 ? 0.03 nM) and exhibits a significantly extended plasma half-life (>6 days) in mice relative to hEPO (?4 h). Mice treated with the Ab-hEPO fusion protein show sustained elevated hematocrit for more than two weeks. This work demonstrates the utility of BLV1H12 CDR3 fusions as a novel approach for generating potent polypeptides with enhanced pharmacological properties.

SUBMITTER: Zhang Y

PROVIDER: S-EPMC3800483 | biostudies-literature | 2013 Oct

REPOSITORIES: biostudies-literature

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An antibody CDR3-erythropoietin fusion protein.

Zhang Yong Y Wang Danling D Welzel Gus G Wang Ying Y Schultz Peter G PG Wang Feng F

ACS chemical biology 20130814 10

X-ray crystallographic analysis of a bovine antibody (BLV1H12) revealed a unique scaffold in its ultralong heavy chain complementarity determining region 3 (CDR3H) that folds into a solvent exposed, antiparallel β-stranded "stalk" fused with a disulfide cross-linked "knob" domain. This unusual variable region motif provides a novel approach for generating chimeric antibodies with novel activities. Toward this end, human erythropoietin (hEPO) was substituted for the "knob" domain in this antibody ...[more]

PMID: 23941200

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