Shark Variable New Antigen Receptor (VNAR) Single Domain Antibody Fragments: Stability and Diagnostic Applications
Shark Variable New Antigen Receptor (VNAR) Single Domain Antibody Fragments: Stability and Diagnostic Applications
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DOI：
10.3390/antib2010066
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发表时间：
2013-03-01
期刊：
影响因子：
4.7
通讯作者：
Foley, Michael
中科院分区：
文献类型：
--
作者：
Griffiths, Katherine;Dolezal, Olan;Foley, Michael
来源于鲨鱼免疫球蛋白新抗原受体抗体（IgNARs）的单可变新抗原受体结构域抗体片段（V（NAR）s）代表了一些已知的最小的基于免疫球蛋白的蛋白质支架。作为单结构域，它们表现出有利的尺寸和隐蔽表位识别特性，使它们在许多疾病状态的诊断和治疗中具有吸引力。在这里，我们研究了稳定性的V-NAR域的重点是V（NAR）的家庭特异性顶端膜抗原1（AMA-1）从恶性疟原虫。将V（NAR）与液体、冻干和固定化硝酸纤维素形式的传统单克隆抗体（mAb）进行比较。当在45 ℃下以各种形式维持时，与mAb相比，V（NAR）具有长达四周的稳定性。使用圆二色性光谱，我们证明V-NAR结构域能够在加热至80摄氏度后重新折叠。我们还证明，V-NAR结构域是稳定的，在潜在的体内条件下，如胃酸孵育，但不是蛋白酶丰富的环境中的小鼠胃刮。总之，我们的研究结果表明鲨鱼V-NAR域的适用性，各种诊断平台和相关应用。
The single variable new antigen receptor domain antibody fragments (V(NAR)s) derived from shark immunoglobulin new antigen receptor antibodies (IgNARs) represent some of the smallest known immunoglobulin-based protein scaffolds. As single domains, they demonstrate favorable size and cryptic epitope recognition properties, making them attractive in diagnosis and therapy of numerous disease states. Here, we examine the stability of V-NAR domains with a focus on a family of V(NAR)s specific for apical membrane antigen 1 (AMA-1) from Plasmodium falciparum. The V(NAR)s are compared to traditional monoclonal antibodies (mAbs) in liquid, lyophilized and immobilized nitrocellulose formats. When maintained in various formats at 45 degrees C, V(NAR)s have improved stability compared to mAbs for periods of up to four weeks. Using circular dichroism spectroscopy we demonstrate that V-NAR domains are able to refold following heating to 80 degrees C. We also demonstrate that V-NAR domains are stable during incubation under potential in vivo conditions such as stomach acid, but not to the protease rich environment of murine stomach scrapings. Taken together, our results demonstrate the suitability of shark V-NAR domains for various diagnostic platforms and related applications.