Purification, Characterization and Plasma Half-Life of PEGylated Soluble Recombinant Non-HA-Binding CD44

Abstract

In summary, we have developed a purification protocol for large-scale production of CD44-3MUT and generated a PEGylated form of CD44-3MUT. HA binding domain of CD44(CD44HABD) and its modified non-HA binding form (CD44-3MUT) inhibit angiogenesis and tumor growth in vivo without disturbing HA-binding functions. CD44-3MUT has been PEGylated for use as a new type of anti-angiogenic human drug. PEGylation of CD44-3MUT improved pharmacokinetic properties but retains its functional activity.

CD44 Controls Endothelial Proliferation and Functions as Endogenous Inhibitor of Angiogenesis

“…2 in Online Resource 1). The volume of distribution of CD44 3MUTFc was 18% of total body weight (%TBW), suggesting improved biodistribution compared to untagged CD443MUT protein (1.8% TBW; [20]). We tested the antiangiogenic effects of CD443MUTFc in athymic nude mice.…”

“…We have previously shown that bacterially expressed GST-tagged HA-nonbinding CD44 (CD44-3MUT) inhibits angiogenesis in chick chorioallantoic membrane and subcutaneous tumor xenograft growth in mice (Päll et al, 2004). Untagged and GST-tagged CD44-3MUT display very short serum half-life limiting their potential in vivo use (Pink et al, 2014). Thus, in order to improve in vivo efficacy, we generated CD44-3MUT with a C-terminal fusion of human IgG1 Fc region (CD44-3MUT-Fc).…”

“…Pharmacokinetic characterization of CD44-3MUT-Fc in rats after intravenous administration showed that its serum half-life is 17 min ( Figure S1). Volume of distribution of CD44-3MUT-Fc is 18 % total body weight (%TBW), suggesting improved biodistribution compared to untagged CD44-3MUT protein (1.8 %TBW; Pink et al (2014)).…”