Figure 1C_Heat Map of gH/gL binding screen

Mice were repeatedly immunized with HCMV purified virus or plasmid encoding gH/gL and blood was collected to evaluate the neutralization capacity of mouse sera in epithelial (ARPE-19) and fibroblast (MRC5) cells using reporter virus AD169R (MOI 0.1). Percent infection was quantified using GFP and normalized to virus incubated in the presence of normal mouse serum (NMS). Each serum condition was tested in technical triplicate (n=3). (c) Antibody clones capable of binding gH/gL were identified following hybridoma supernatant incubation with 293ExpiF cells transiently expressing gH/gL. Mean fluorescence intensity (MFI) was determined for each sample and summarized by fusion (n=1).

以纯化的人类巨细胞病毒（HCMV）或编码糖蛋白H/糖蛋白L（gH/gL）的质粒对小鼠进行反复免疫，采集血液样本，采用报告病毒（reporter virus）AD169R（感染复数（Multiplicity of Infection，MOI）为0.1），在上皮细胞（ARPE-19）和成纤维细胞（MRC5）中检测小鼠血清的中和活性。通过绿色荧光蛋白（Green Fluorescent Protein，GFP）对感染百分比进行量化，并以正常小鼠血清（Normal Mouse Serum，NMS）孵育病毒的实验组作为参照完成标准化处理。每个血清实验组均设置三次技术重复检测（n=3）。(c) 将杂交瘤细胞上清与瞬时表达gH/gL的293ExpiF细胞共孵育后，可鉴定出能够结合gH/gL的抗体克隆。对每个样品测定平均荧光强度（Mean Fluorescence Intensity，MFI），并按融合实验进行汇总（n=1）。