利用细菌全细胞作为分子探针分离病原体特异性人源单克隆抗体（hmAbs）。
Isolating Pathogen-Specific Human Monoclonal Antibodies (hmAbs) Using Bacterial Whole Cells as Molecular Probes.
机构信息
Section of Paediatric Infectious Disease, Department of Infectious Disease, Imperial College London, London, UK.
Flow Cytometry Core Facility, National Heart and Lung Institute, Imperial College London, London, UK.
出版信息
Methods Mol Biol. 2021;2183:9-18. doi: 10.1007/978-1-0716-0795-4_2.
The immunoglobulin capture assay (ICA) enables the enrichment for pathogen-specific plasmablasts from individuals with a confirmed adaptive immune response to vaccination or disseminated infection. Only single recombinant antigens have been used previously as probes in this ICA and it was unclear whether the method was applicable to complex probes such as whole bacterial cells. Here, we describe the enrichment of plasmablasts specific for polysaccharide and protein antigens of both Streptococcus pneumoniae and Neisseria meningitidis using whole formalin-fixed bacterial cells as probes. The modified ICA protocol described here allowed for a pathogen-specific hmAb cloning efficiency of >80%.
免疫球蛋白捕获测定法（ICA）可从接种疫苗或发生全身性感染后具有明确适应性免疫反应的个体中富集针对病原体的浆母细胞。此前，该 ICA 仅使用单一重组抗原作为探针，尚不清楚该方法是否适用于全细菌细胞等复杂探针。在此，我们描述了使用全福尔马林固定细菌细胞作为探针，从肺炎链球菌和脑膜炎奈瑟菌的多糖和蛋白抗原中富集针对特定抗原的浆母细胞。本文所述改良 ICA 方案可使病原体特异性人源化单抗的克隆效率＞80%。
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