首页>
外国专利>
Method for the identification and isolation of binding polypeptides from combinatorial libraries of proteins having the scaffold structure of C-type lectin-like domains
Method for the identification and isolation of binding polypeptides from combinatorial libraries of proteins having the scaffold structure of C-type lectin-like domains
展开▼
机译:从具有C型凝集素样结构域的支架结构的蛋白质的组合文库中鉴定和分离结合多肽的方法
展开▼
页面导航
摘要
著录项
相似文献
摘要
A novel family of protein libraries comprising CTLDs (C-type Lectin-Like Domains) in which internal polypeptide loop-regions lining the ligand binding sites in CTLDs have been replaced with ensembles of completely or partially randomised polypeptide segments. Tetranectin CTLDs were chosen as framework for the preferred embodiment of the invention; and versatile phagemid vectors useful in the generation and manipulation of human and murine tetranectin CTLD libraries are disclosed as part of this invention. Tetranectin CTLDs in monomeric as well as in trimeric form are efficiently displayed as gene III fusions in fully functional form by the recombinant fd phage display vector. CTLD derivatives with affinity for new ligands may readily be isolated from libraries of vectors displaying CTLDs, in which loop-regions have been randomised, using one or more rounds of enrichment by screening or selection followed by amplification of the enriched subpopulation in each round. The efficiency with which protein products containing CTLDs with new binding properties can be produced, e.g. by bacterial expression and in vitro refolding, in mono-, tri-, or multimeric formats provides important advantages in terms of simplicity, cost and efficiency of generation, production and diagnostic or therapeutic applications in comparison to recombinant antibody derivatives.
展开▼