Mena Jimmy A, Ramírez Octavio T, Palomares Laura A
Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal. 510-3, Cuernavaca, Morelos CP. 62250, México.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Sep 25;824(1-2):267-76. doi: 10.1016/j.jchromb.2005.07.034.
There is a lack of accurate and practical methods that require only small amounts of sample for quantifying virus-like particles (VLP). In this work, gel permeation (GP) HPLC was used to quantify double-layered rotavirus-like particles (dlRLP) produced in insect cells. The proposed methodology utilized two columns in series (pore sizes of 200 and 50 nm) and had a high precision (relative standard deviation below 5%). GP-HPLC not only allowed the routine quantification of dlRLP, but also of assembly intermediaries and other viral structures present in the samples. For the first time, kinetics of dlRLP accumulation could be followed. This methodology is valuable for designing new production processes and for optimizing dlRLP monitoring.
目前缺乏仅需少量样品就能定量病毒样颗粒（VLP）的准确实用方法。在本研究中，采用凝胶渗透（GP）高效液相色谱法对昆虫细胞中产生的双层轮状病毒样颗粒（dlRLP）进行定量。所提出的方法使用两根串联柱（孔径分别为200和50纳米），具有很高的精密度（相对标准偏差低于5%）。GP-HPLC不仅能对dlRLP进行常规定量，还能对样品中存在的组装中间体和其他病毒结构进行定量。首次能够跟踪dlRLP的积累动力学。该方法对于设计新的生产工艺和优化dlRLP监测具有重要价值。
