Milford Hospital and 1Milford Molecular Laboratory, 2044 Bridgeport Avenue, Milford, CT 06460, USA.
Curr Med Chem. 2014 Mar;21(7):932-40. doi: 10.2174/0929867321999140102110933.
Gardasil® is a quadrivalent human papillomavirus (HPV) protein-based vaccine containing genotype-specific L1 capsid proteins of HPV-16, HPV-18, HPV-6 and HPV-11 in the form of virus-like-particles (VLPs) as the active ingredient. The VLPs are produced by a DNA recombinant technology. It is uncertain if the residual HPV L1 gene DNA fragments in the vaccine products are considered contaminants or excipients of the Gardasil® vaccine. Because naked viral DNA fragments, if present in the vaccine, may bind to the insoluble amorphous aluminum hydroxyphosphate sulfate (AAHS) adjuvant which may help deliver the foreign DNA into macrophages, causing unintended pathophysiologic effects, experiments were undertaken to develop tests for HPV L1 gene DNA fragments in the final products of Gardasil® by polymerase chain reaction (PCR) and direct DNA sequencing. The results showed that while the HPV-11 and HPV-18 L1 gene DNA fragments in Gardasil® were readily amplified by the common GP6/MY11 degenerate consensus primers, the HPV-16 L1 gene DNA may need specially designed non-degenerate PCR primers for amplification at different regions of the L1 gene and different stringency conditions for detection. These variable melting profiles of HPV DNA in the insoluble fraction of the Gardasil® vaccine suggest that the HPV DNA fragments are firmly bound to the aluminum AAHS adjuvant. All methods developed for detecting residual HPV DNA in the vaccine Gardasil® for quality assurance must take into consideration the variable melting profiles of the DNA to avoid false negative results.
佳达修®是一种四价人乳头瘤病毒（HPV）蛋白疫苗，包含 HPV-16、HPV-18、HPV-6 和 HPV-11 型特异性 L1 衣壳蛋白，以病毒样颗粒（VLPs）的形式作为活性成分。VLPs 是通过 DNA 重组技术生产的。疫苗产品中是否存在 HPV L1 基因 DNA 片段被认为是污染物或佳达修®疫苗的赋形剂尚不确定。因为如果疫苗中存在裸露的病毒 DNA 片段，可能会与不溶性无定形羟基磷灰石硫酸铝（AAHS）佐剂结合，这可能有助于将外源 DNA 递送至巨噬细胞，引起意想不到的病理生理效应，因此进行了实验以开发聚合酶链反应（PCR）和直接 DNA 测序用于检测佳达修®最终产品中 HPV L1 基因 DNA 片段的方法。结果表明，虽然佳达修®中的 HPV-11 和 HPV-18 L1 基因 DNA 片段可以用常见的 GP6/MY11 简并共识引物轻易扩增，但 HPV-16 L1 基因 DNA 可能需要专门设计的非简并 PCR 引物来扩增 L1 基因的不同区域和不同的检测严格性条件。HPV DNA 在佳达修®疫苗不溶性部分的这些可变熔解曲线表明 HPV DNA 片段与铝 AAHS 佐剂牢固结合。用于检测疫苗佳达修®中残留 HPV DNA 的所有方法都必须考虑 DNA 的可变熔解曲线，以避免假阴性结果。
