公开(公告)号：US20070154901A1

公开(公告)日：2007-07-05

申请号：US11452434

申请日：2006-06-14

申请人： Hans Thogersen ,  Michael Etzerodt ,  Thor Holtet ,  Niels Graversen ,  Jette Kastrup ,  Bettina Nielsen ,  Ingrid Larsen

发明人： Hans Thogersen ,  Michael Etzerodt ,  Thor Holtet ,  Niels Graversen ,  Jette Kastrup ,  Bettina Nielsen ,  Ingrid Larsen

IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P21/06 ,  C07K14/47

CPC分类号： C12N15/62 ,  A61K38/00 ,  A61K48/00 ,  C07K14/4726 ,  C07K16/00 ,  C07K2319/00 ,  C07K2319/41 ,  C07K2319/50 ,  C07K2319/73 ,  C07K2319/95

摘要： The present invention relates to the design of trimeric polypeptides using polypeptide structural elements derived from the tetranectin protein family, and their use in rational de novo design and production of multi-functional molecules including the application of the multi-functional molecules in protein library technology, such as phage display technology, diagnostic and therapeutic systems, such as human gene therapy and imaging. The trimeric polypeptides being constructed as a monomer polypeptide construct comprising at least one tetranectin trimerising structural element (TTSE) which is covalently linked to at least one heterologous moiety, said TTSE being capable of forming a stable complex with two other TTSEs; or as an oligomer which is comprised of two monomer polypeptide constructs as mentioned above, and which comprises three TTSEs or a multiplum of three TTSEs, or which is comprised of three monomer polypeptide constructs.

摘要翻译： 本发明涉及使用衍生自tetranectin蛋白家族的多肽结构元件的三聚体多肽的设计及其在合理的从头设计和生产多功能分子的用途，包括在蛋白质文库技术中应用多功能分子， 如噬菌体展示技术，诊断和治疗系统，如人类基因治疗和成像。 三聚体多肽被构建为包含至少一种与至少一个异源部分共价连接的四联蛋白三聚结构元件（TTSE）的单体多肽构建体，所述TTSE能够与两个其它TTSE形成稳定的复合物; 或作为如上所述由两个单体多肽构建体组成的寡聚体，并且其包含三个TTSE或三个TTSE的多重，或由三个单体多肽构建体组成。

公开(公告)号：US20070275393A1

公开(公告)日：2007-11-29

申请号：US11633040

申请日：2006-12-04

申请人： Michael Etzerodt ,  Thor Holtet ,  Niels Graversen ,  Hans Thogersen

发明人： Michael Etzerodt ,  Thor Holtet ,  Niels Graversen ,  Hans Thogersen

IPC分类号： C12Q1/68 ,  C07K14/47 ,  C12P21/00

CPC分类号： C12N15/1044 ,  C07K14/4726 ,  C40B40/02 ,  C40B40/08 ,  C40B50/06

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.

摘要翻译： 包含CTLD（C型凝集素样结构域）的蛋白质文库的新家族，其中在CTLD中配体结合位点衬里的内部多肽环区域被完全或部分随机化的多肽片段的整体替代。 选择Tetranectin CTLD作为本发明优选实施方案的框架; 并且用于生成和操纵人和鼠四联蛋白CTLD文库的多功能噬菌粒载体作为本发明的一部分被公开。 通过重组fd噬菌体展示载体，单体和三聚体形式的Tetranectin CTLD有效地显示为完全功能形式的基因III融合体。 对新配体具有亲和力的CTLD衍生物可以容易地从显示CTLD的载体文库中分离，其中环区已被随机化，使用一轮或多轮浓缩通过筛选或选择，随后在每轮中扩增富集的亚群。 可以生产含有具有新结合特性的CTLD的蛋白质产物的效率，例如， 通过细菌表达和体外重折叠，单 - ，三 - 或多聚体形式在与重组抗体衍生物相比的简单性，成本和生产效率，生产和诊断或治疗应用方面提供了重要的优点。