Self-Assembly of Antibodies by Chemical Induction
Self-Assembly of Antibodies by Chemical Induction
复制标题
DOI：
10.1002/anie.200803507
复制
发表时间：
2008-01-01
影响因子：
16.6
通讯作者：
Wagner, Carston R.
中科院分区：
文献类型：
--
作者：
Li, Qing;Hapka, David;Wagner, Carston R.
克隆和表达单克隆抗体的可变（VH和VL）和恒定（CH和CL）结构域的能力使得构建基因工程抗体成为可能。[1-3]抗体片段，例如Fab，已经融合到自组装蛋白结构域，例如转录因子的亮氨酸拉链区或IgG的CL和CH 1结构域。[4，5]重组抗体，称为单链抗体（scFv），已经通过经由肽接头融合抗体可变区（Fv）的VH和VL结构域来构建。这一进展导致了用于制备由单个多肽组成的相应融合相关物、双抗体（二价）和tandabs（四价）的多种方法的发展。[6-10]虽然目前制备重组抗体的方法具有多种优点，但一般来说，如果不求助于蛋白质解折叠所需的条件，就不可能控制它们的组装和拆卸。由于工程化抗体的分子量和尺寸是其体内亲合力、生物分布和药代动力学的重要决定因素，因此允许通过化学控制的组装和拆卸暂时改变这些参数的方法原则上将增强重组抗体的治疗和诊断效用[11，12]。我们开发了一种通过用双甲氨蝶呤（Bis-MTX）化学诱导制备二氢叶酸还原酶融合蛋白（DHFR 2）的自组装蛋白质大环低聚物或纳米环的方案。直径从7到30 nm的自组装纳米环可以很容易地分别从2到8个DHFR 2单体制备。[13]纳米环的大小被发现是由特定的长度和组成的肽插入之间的DHF。例如，当DHFR 2之间的接头肽是13个氨基酸的柔性接头时，
The ability to clone and express the variable (VH & VL) and constant (CH & CL) domains of monoclonal antibodies has made the construction of genetically engineered antibodies possible.[1-3] Antibody fragments, such as Fabs, have been fused to self-assembling protein domains such as leucine zipper regions of the transcription factors or to the CL and CH1 domains of IgG.[4, 5] Recombinant antibodies, referred to as single chain antibodies (scFv's), have been constructed by fusing the VH and VL domains of an antibody variable region (Fv) via a peptide linker. This advance resulted in the development of a variety of approaches for the preparation of the corresponding fusion relatives, diabodies (bivalent) and tandabs (tetravalent) consisting of a single polypeptide.[6-10] Although the current methods of preparing recombinant antibodies have a variety of advantages, in general control over their assembly and disassembly is not possible without resorting to conditions necessary for protein unfolding. Since the molecular weight and dimensions of engineered antibodies is a significant determinant of their in vivo avidity, biodistribution and pharmacokinetics, a method allowing these parameters to be temporally altered by chemically controlled assembly and disassembly would in principle enhance the therapeutic and diagnostc utility of recombinant antibodies [11, 12].Recently, we have developed a protocol for the preparation of self-assembling protein macrocyclic oligomers, or nanorings, of dihydrofolate reductase fusion proteins (DHFR2) by chemical induction with bis-methotrexate (Bis-MTX). Self-assembling nanorings ranging in diameter from 7 to 30 nm could easily be prepared from two to eight DHFR2 monomers, respectively.[13] The nanoring size was found to be governed by the specific length and composition of the peptide inserted between the DHFRs. For example, when the linker peptide between the DHFR2 was a 13 amino acid flexible linker, dimer formation was