Chai Zheng, Samulski R Jude, Li Chengwen
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Bio Protoc. 2018 May 5;8(9). doi: 10.21769/BioProtoc.2841.
Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These AAV chimeric capsid mutants enhanced Nab evasion from patient serum with a high muscle transduction efficacy. In this protocol, we describe the procedures for selection of the Nab-escaped AAV chimeric capsid, including isolation and characterization of Nab-escaping AAV mutants in mice muscle.
中和抗体（Nabs）是腺相关病毒（AAV）载体基因治疗临床试验中的一个主要挑战，因为中和抗体能够抑制患者体内的AAV转导。我们通过给予AAV随机文库和患者血清的混合物，成功地在小鼠中分离出了几种新型的逃避中和抗体的AAV嵌合衣壳。这些AAV嵌合衣壳突变体增强了对患者血清中中和抗体的逃避能力，并具有高肌肉转导效率。在本方案中，我们描述了选择逃避中和抗体的AAV嵌合衣壳的程序，包括在小鼠肌肉中分离和鉴定逃避中和抗体的AAV突变体。
