Isolating Pathogen-Specific Human Monoclonal Antibodies (hmAbs) Using Bacterial Whole Cells as Molecular Probes
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%.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:2183 |
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| Enthalten in: |
Methods in molecular biology (Clifton, N.J.) - 2183(2021) vom: 22., Seite 9-18 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Siris, Sara [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 22.03.2021 Date Revised 22.03.2021 published: Print Citation Status MEDLINE |
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| doi: |
10.1007/978-1-0716-0795-4_2 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM315304669 |
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| 520 | |a 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% | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Bacteria | |
| 650 | 4 | |a Immunoglobulin capture assay | |
| 650 | 4 | |a Pathogen-specific plasmablasts | |
| 650 | 4 | |a Vaccine antigen discovery | |
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| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a Antigens, Bacterial |2 NLM | |
| 650 | 7 | |a Immunoglobulin G |2 NLM | |
| 650 | 7 | |a Molecular Probes |2 NLM | |
| 700 | 1 | |a Gladstone, Camilla A |e verfasserin |4 aut | |
| 700 | 1 | |a Guo, Yanping |e verfasserin |4 aut | |
| 700 | 1 | |a Pinder, Christopher L |e verfasserin |4 aut | |
| 700 | 1 | |a Shattock, Robin J |e verfasserin |4 aut | |
| 700 | 1 | |a McKay, Paul F |e verfasserin |4 aut | |
| 700 | 1 | |a Langford, Paul R |e verfasserin |4 aut | |
| 700 | 1 | |a Bidmos, Fadil A |e verfasserin |4 aut | |
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