Single-chain Fv with manifold N-glycans as bifunctional scaffolds for immunomolecules |
| |
Authors: | Wang M; Lee LS; Nepomich A; Yang JD; Conover C; Whitlow M; Filpula D |
| |
Affiliation: | Enzon, Incorporated, Piscataway, NJ 08854-3969, USA. |
| |
Abstract: | Unlike natural antibodies, single-chain Fv (sFv) proteins normally lack
asparagine-linked glycosylation. Many designed immunoconjugates and other
therapeutics currently employ the advantageous conjugation chemistry or
targeting properties provided by the glycoprotein oligosaccharide domain.
sFv proteins with engineered N-glycan designs were evaluated in Pichia
pastoris for glycosylation efficiency, expression level, oligosaccharide
chain length and composition, and affinity. In contrast to nearly all
natural glycoproteins, the engineered attachment of N-glycans conveniently
near the polypeptide C- terminus was found to produce the optimal results.
Furthermore, the percentage modification and chain length of the attached
mannose chains were controllable by the use of tandem and overlapping
Asn-X-Thr tripeptide sites. The glycosylated sFv mannose chains could be
effectively conjugated to polyethylene glycol and the resulting conjugate
displayed a 10-fold increased circulating life in mice. The potential to
control polymer:sFv or drug:sFv molar ratios by site- specific conjugation
may substantially improve the therapeutic efficacy of these minimal
antigen-binding molecules.
|
| |
Keywords: | |
本文献已被 Oxford 等数据库收录! |
|