FACS analyses of purified RhD-specific IgG antibodies

unstained.001<br>FACS analysis with Rhesus D (RhD)-positive erythrocytes to verify the enrichment of RhD-specific IgGs from total IgG of Rhophylac. The cells in this file are unstained. The cells were gated on the main erythrocyte population in a FSC/SSC blot to exclude fragments and aggregates, analyzed in an anti-human IgG (APC) histogram and shown in the overlay histogram in Supplementary Figure 1 in the main text. only_secondary_antibody.004<br>FACS analysis with RhD-positive erythrocytes to verify the enrichment of RhD-specific IgGs from total IgG of Rhophylac.<br>The cells in this file are only stained with a secondary APC-coupled anti-human IgG antibody. The cells were gated on the main erythrocyte population in a FSC/SSC blot to exclude fragments and aggregates, analyzed in an anti-human IgG (APC) histogram and shown in the overlay histogram in Supplementary Figure 1 in the main text. 260_µg_per_ml_total_IgG_from_Rhophylac.002<br>FACS analysis with RhD-positive erythrocytes to verify the enrichment of RhD-specific IgGs from total IgG of Rhophylac.<br>The cells in this file are stained with 260 µg/ml total IgG from Rhophylac and subsequently with a secondary APC-coupled anti-human IgG antibody.The cells were gated on the main erythrocyte population in a FSC/SSC blot to exclude fragments and aggregates, analyzed in an anti-human IgG (APC) histogram and shown in the overlay histogram in Supplementary Figure 1 in the main text. 2µg_per_ml_purified_RhD_specific_IgG(from_purification_A).005<br>FACS analysis with RhD-positive erythrocytes to verify the enrichment of RhD-specific IgGs from total IgG of Rhophylac.<br>The cells in this file are stained with 2 µg/ml purified RhD-specific IgG from purification A and subsequently with a secondary APC-coupled anti-human IgG antibody. The cells were gated on the main erythrocyte population in a FSC/SSC blot to exclude fragments and aggregates, analyzed in an anti-human IgG (APC) histogram and shown in the overlay histogram in Supplementary Figure 1 in the main text. 2µg_per_ml_purified_RhD_specific_IgG(from_purification_B).007<br>FACS analysis with RhD-positive erythrocytes to verify the enrichment of RhD-specific IgGs from total IgG of Rhophylac.<br>The cells in this file are stained with 2 µg/ml purified RhD-specific IgG from purification B and subsequently with a secondary APC-coupled anti-human IgG antibody.<br>These cells are not shown in the overlay histogram in Supplementary Figure 1 in the main text. However, the staining looks like the staining from the file before. 2_µg_per_ml_total_IgG_from_Rhophylac.003<br>FACS analysis with RhD-positive erythrocytes to verify the enrichment of RhD-specific IgGs from total IgG of Rhophylac.<br>The cells in this file are stained with 2 µg/ml total IgG from Rhophylac and subsequently with a secondary APC-coupled anti-human IgG antibody. The cells were gated on the main erythrocyte population in a FSC/SSC blot to exclude fragments and aggregates, analyzed in an anti-human IgG (APC) histogram and shown in the overlay histogram in Supplementary Figure 1 in the main text.

未染色组.001
采用RhD阳性红细胞开展荧光激活细胞分选术（Fluorescence-Activated Cell Sorting，FACS）分析，以验证从Rhophylac总免疫球蛋白G（Immunoglobulin G，IgG）中富集RhD特异性IgG的效果。本文件所包含的细胞未进行染色。通过前向散射光（Forward Scatter，FSC）/侧向散射光（Side Scatter，SSC）散点图对红细胞主群体设门，以排除细胞碎片与聚集体；随后在别藻蓝蛋白（Allophycocyanin，APC）标记抗人IgG直方图中进行数据分析，结果展示于正文补充图1的叠加直方图中。
仅二抗组.004
采用RhD阳性红细胞开展FACS分析，以验证从Rhophylac总IgG中富集RhD特异性IgG的效果。本文件所包含的细胞仅使用别藻蓝蛋白（APC）偶联抗人IgG二抗进行染色。通过FSC/SSC散点图对红细胞主群体设门，以排除细胞碎片与聚集体；随后在APC标记抗人IgG直方图中进行数据分析，结果展示于正文补充图1的叠加直方图中。
260μg/ml Rhophylac总IgG组.002
采用RhD阳性红细胞开展FACS分析，以验证从Rhophylac总IgG中富集RhD特异性IgG的效果。本文件所包含的细胞先用260μg/mL的Rhophylac总IgG染色，随后使用别藻蓝蛋白（APC）偶联抗人IgG二抗进行标记。通过FSC/SSC散点图对红细胞主群体设门，以排除细胞碎片与聚集体；随后在APC标记抗人IgG直方图中进行数据分析，结果展示于正文补充图1的叠加直方图中。
2μg/ml 纯化批次A来源RhD特异性IgG组.005
采用RhD阳性红细胞开展FACS分析，以验证从Rhophylac总IgG中富集RhD特异性IgG的效果。本文件所包含的细胞先用2μg/mL的纯化批次A来源的RhD特异性IgG染色，随后使用别藻蓝蛋白（APC）偶联抗人IgG二抗进行标记。通过FSC/SSC散点图对红细胞主群体设门，以排除细胞碎片与聚集体；随后在APC标记抗人IgG直方图中进行数据分析，结果展示于正文补充图1的叠加直方图中。
2μg/ml 纯化批次B来源RhD特异性IgG组.007
采用RhD阳性红细胞开展FACS分析，以验证从Rhophylac总IgG中富集RhD特异性IgG的效果。本文件所包含的细胞先用2μg/mL的纯化批次B来源的RhD特异性IgG染色，随后使用别藻蓝蛋白（APC）偶联抗人IgG二抗进行标记。本样本未展示于正文补充图1的叠加直方图中，但染色效果与前一样本一致。
2μg/ml Rhophylac总IgG组.003
采用RhD阳性红细胞开展FACS分析，以验证从Rhophylac总IgG中富集RhD特异性IgG的效果。本文件所包含的细胞先用2μg/mL的Rhophylac总IgG染色，随后使用别藻蓝蛋白（APC）偶联抗人IgG二抗进行标记。通过FSC/SSC散点图对红细胞主群体设门，以排除细胞碎片与聚集体；随后在APC标记抗人IgG直方图中进行数据分析，结果展示于正文补充图1的叠加直方图中。