Bender Michael F, Li Yile, Ivleva Vera B, Gowetski Daniel B, Paula Lei Q
Vaccine Production Program Laboratory, Vaccine Research Center, National Institute of Allergy and Infectious Disease, United States.
Vaccine Production Program Laboratory, Vaccine Research Center, National Institute of Allergy and Infectious Disease, United States.
Vaccine. 2021 Jun 23;39(28):3650-3654. doi: 10.1016/j.vaccine.2021.05.048. Epub 2021 May 28.
High Performance Size-Exclusion Chromatography coupled with Multi-Angle Light Scattering detection (HPSEC-MALS) is an important tool to provide a reliable molecular weight measurement for a large complex biomolecule. A recent HIV-1 soluble envelope trimer vaccine candidate, BG505 DS-SOSIP.664, is among the most glycosylated proteins to enter a clinical trial to date, and determination of its protein and glycan molecular weight is one of the key attributes in pre-clinical characterization. However, protein and glycans possess disparate dndcvalues making molecular weight measurement inaccurate in conventional SEC-MALS. To overcome these challenges, a simple mathematically guided experiment was explored, and a composite dndcvalue was established by utilizing protein and glycan mass contributions for the HIV-1 envelope trimer. This establishment was further verified by an orthogonal mass spectrometry analysis. This innovative, simple, and quick analytical approach can be applied broadly to measuring the molecular weight of various composite molecular structures, such as complex glycoconjugates.
高效尺寸排阻色谱联用多角度光散射检测技术（HPSEC-MALS）是为大型复杂生物分子提供可靠分子量测量的重要工具。一种近期的HIV-1可溶性包膜三聚体候选疫苗BG505 DS-SOSIP.664是迄今为止进入临床试验的糖基化程度最高的蛋白质之一，确定其蛋白质和聚糖分子量是临床前表征的关键特性之一。然而，蛋白质和聚糖具有不同的dn/dc值，使得传统SEC-MALS中的分子量测量不准确。为了克服这些挑战，探索了一个简单的数学指导实验，并利用HIV-1包膜三聚体的蛋白质和聚糖质量贡献建立了复合dn/dc值。通过正交质谱分析进一步验证了这一建立过程。这种创新、简单且快速的分析方法可广泛应用于测量各种复合分子结构的分子量，如复杂糖缀合物。
