KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Peroxidase-Conjugated Goat Affinity Purified Antibody to Mouse IgG (Whole Molecule) | MP Biomedical | Cat# 0855563 |
| Peroxidase AffiniPure F(ab’)2 Fragment Goat Anti-Human IgG | Jackson ImmunoResearch Laboratories | Cat# 109-036-098 |
| Mouse anti-NP antibody (IC5-1B7) | BEI Resources | Cat# NR-43899 |
| Monoclonal Anti-Human Influenza A (H3N2) (Clone F49) | Takara Bio | Cat# M146 |
| Monoclonal Anti-Human Influenza A (H1N1, H2N2) (Clone C179) | Takara Bio | Cat# M145 |
| 008-10053 1G05 | This study | N/A |
| 008-10053 5E04 | This study | N/A |
| 008-10053 5G01 | This study | N/A |
| 008-10053 6C05 | This study | N/A |
| 009-10061 2A05 | This study | N/A |
| 009-10061 2C06 | This study | N/A |
| 009-10061 3B06 | This study | N/A |
| 011-10069 2C01 | This study | N/A |
| 011-10069 3E06 | This study | N/A |
| 011-10069 5C01 | This study | N/A |
| 011-10069 5D01 | This study | N/A |
| 011-10069 5G01 | This study | N/A |
| 011-10069 5G04 | This study | N/A |
| 013-10078 3G01 | This study | N/A |
| 017-10116 5B03 | This study | N/A |
| 017-10116 5D02 | This study | N/A |
| 017-10116 5G02 | This study | N/A |
| 019-10117 1B02 | This study | N/A |
| 019-10117 3A06 | This study | N/A |
| 019-10117 3C06 | This study | N/A |
| 019-10117 3F01 | This study | N/A |
| 019-10117 3F02 | This study | N/A |
| 024-10128 3C04 | This study | N/A |
| 024-10128 3F04 | This study | N/A |
| 028-10134 4F03 | This study | N/A |
| 030-121509 3B01 | This study | N/A |
| 034-10040 4C01 | This study | N/A |
| 034-10040 4E01 | This study | N/A |
| 034-10040 4F02 | This study | N/A |
| 034-100809 3E01 | This study | N/A |
| 037-10036 5A01 | This study | N/A |
| 041-10047 1C04 | This study | N/A |
| 042-10065 2B03 | This study | N/A |
| Bacterial and Virus Strains | ||
| A/Hong Kong/1/1968 H3N2 | BEI | NR-346 |
| A/Scotland/840/1974 H3N2 | BEI | NR-3662 |
| A/Mississippi/1/1985 H3N2 | BEI | NR-3502 |
| A/Johannesburg/33/1994 H3N2 | BEI | NR-3656 |
| A/Wisconsin/67/2005 H3N2 | IRR | FR-397 |
| A/Victoria/210/2009 H3N2 (X-187, GenBank HQ378745) and point mutants | Reverse-genetics derived, this study | N/A |
| A/Perth/16/2009 H3N2 and point mutants | Reverse-genetics derived, this study and (Lee et al., 2018) | N/A |
| A/Texas/50/2012 H3N2 | Reverse-genetics derived, (Chambers et al., 2015) | N/A |
| A/Switzerland/9715293/2013 H3N2 (GISAID EPI543062) | Reverse-genetics derived | N/A |
| A/Hong Kong/4801/2014 H3N2 (GenBank KT838266.1) | Reverse-genetics derived, this study | N/A |
| A/California/07/2009 H1N1 (GenBank NC_026433.1) with D225G substitution | Reverse-genetics derived, this study | N/A |
| A/Perth/16/2009 mutational library | (Lee et al., 2018) | N/A |
| Biological Samples | ||
| Human serum samples before and after 2010-2011 vaccination | This study | N/A |
| Human serum samples before and after 2015-2016 vaccination | This study | N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| TrueBlue TMB Peroxidase Substrate | KPL-Seracare | Cat# 5510-0030 |
| SureBlue TMB Peroxidase Substrate | KPL-Seracare | Cat# 5120-0077 |
| Modified Eagle Medium (MEM) | Corning Cellgro | Cat# MT10-010-CM |
| Dulbecco’s Modified Eagle Medium (DMEM) | Corning Cellgro | Cat# MT10-013-CM |
| FreeStyle 293 Expression Medium | Thermo Fisher | Cat# 12338026 |
| Fetal Bovine Serum | Sigma-Aldrich | Cat# F0926-100ML |
| HEPES buffer | Thermo Fisher | Cat# 25-060-CI |
| Gentamicin | Thermo Fisher | Cat# 15750060 |
| Trypsin, TPCK Treated | Thermo Fisher | Cat# 20233 |
| Bovine Serum Albumin | Sigma-Aldrich | Cat# A8022-500G |
| Hydrochloric acid 37% | Sigma-Aldrich | Cat# 258148-500ML |
| Receptor-destroying enzyme II | VWR | Cat# 10753-482 |
| Tween 20 | Thermo Fisher | Cat# 9005-64-5 |
| Methyl cellulose | Sigma-Aldrich | Cat# M0512 |
| Paraformaldehyde 32% solution | Electron Microscopy Sciences | Cat# 15714-S |
| Triton X-100 | Sigma-Aldrich | Cat# 10789704001 |
| Fat-free milk, dry powder | Dot Scientific, Inc | Cat# DSM17200-1000 |
| 293fectin Transfection Reagent | Thermo Fisher | Cat# 12347019 |
| Ni-NTA Agarose | QIAGEN | Cat# 30210 |
| A/Hong Kong/1/1968 recombinant HA | This study, from baculovirus | N/A |
| A/Perth/16/2009 recombinant HA | This study, from baculovirus | N/A |
| A/Victoria/210/2009 WT recombinant HA | This study, from 293F mammalian expression | N/A |
| A/Victoria/210/2009 Y98F recombinant HA | This study, from 293F mammalian expression | N/A |
| A/Hong Kong/4801/2014 WT recombinant HA | This study, from 293F mammalian expression | N/A |
| A/Hong Kong/4801/2014 Y98F recombinant HA | This study, from 293F mammalian expression | N/A |
| Millipore Protein A/G Magnetic beads | EMD Millipore | Cat # LSKMAGAG10 |
| Critical Commercial Assays | ||
| Quikchange II XL Mutagenesis Kit | Agilent Technologies | Cat# 200522 |
| Deposited Data | ||
| HA mutant libraries selected by monoclonal antibodies | This paper | SAMN10183083 |
| HA monoclonal antibody selection controls | This paper | SAMN10183146 |
| Human Ig-VH and IgVL sequences | GenBank | MH205211, MH205413, KM603994, and MN483379-MN483441 |
| Experimental Models: Cell Lines | ||
| Freestyle 293F | Thermo Fisher | Cat# R79007 |
| 293T | NIH | N/A |
| MDCK | NIH | N/A |
| MDCK-SIAT1 | NIH | N/A |
| Recombinant DNA | ||
| Plasmid: HIV gag (pTR994) | University of Georgia | N/A |
| Plasmid: A/Puerto Rico/1934 NA (VRC9776) | NIH | N/A |
| Plasmid: HAT protease (HAT in pCAGGS) | Doms lab, Penn | N/A |
| Plasmid: influenza reverse genetics plasmids | NIH | N/A |
| Plasmid: A/Victoria/210/2009 HA WT X-187 (GenBank HQ378745) and point mutants (for VLP production) | This study | N/A |
| Plasmid: A/Hong Kong/4801/2014 HA WT (GenBank KT838266.1) and point mutants (for absorption assays) | This study | N/A |
| 008-10053 1G05_Vκ | This study | N/A |
| 008-10053 5E04_Vκ | This study | N/A |
| 008-10053 5G01_Vκ | This study | N/A |
| 008-10053 6C05_Vκ | This study | N/A |
| 009-10061 2A05_Vλ | This study | N/A |
| 009-10061 2C06_Vλ | This study | N/A |
| 009-10061 3B06_VK | This study | N/A |
| 011-10069 2C01_Vλ | This study | N/A |
| 011-10069 3E06_Vλ | This study | N/A |
| 011-10069 5C01_Vλ | This study | N/A |
| 011-10069 5D01_Vλ | This study | N/A |
| 011-10069 5G01_Vλ | This study | N/A |
| 011-10069 5G04_Vλ | This study | N/A |
| 013-10078 3G01_Vλ | This study | N/A |
| 017-10116 5B03_Vκ | This study | N/A |
| 017-10116 5D02_Vκ | This study | N/A |
| 017-10116 5G02_Vκ | This study | N/A |
| 019-10117 1B02_Vκ | This study | N/A |
| 019-10117 3A06_Vλ | This study | N/A |
| 019-10117 3C06_Vλ | This study | N/A |
| 019-10117 3F01_Vλ | This study | N/A |
| 019-10117 3F02_Vλ | This study | N/A |
| 024-10128 3C04_Vλ | This study | N/A |
| 024-10128 3F04_Vλ | This study | N/A |
| 028-10134 4F03_Vκ | This study | N/A |
| 030-121509 3B01_Vκ | This study | N/A |
| 034-10040 4C01_Vκ | This study | N/A |
| 034-10040 4E01_Vλ | This study | N/A |
| 034-10040 4F02_Vκ | This study | N/A |
| 034-100809 3E01_Vκ | This study | N/A |
| 037-10036 5A01_Vλ | This study | N/A |
| 041-10047 1C04_Vλ | This study | N/A |
| 042-10065 2B03_Vλ | This study | N/A |
| 008-10053 1G05_IgG1 | This study | N/A |
| 008-10053 5E04_IgG1 | This study | N/A |
| 008-10053 5G01_IgG1 | This study | N/A |
| 008-10053 6C05_IgG1 | This study | N/A |
| 009-10061 2A05_IgG1 | This study | N/A |
| 009-10061 2C06_IgG1 | This study | N/A |
| 009-10061 3B06_IgG1 | This study | N/A |
| 011-10069 2C01_IgG1 | This study | N/A |
| 011-10069 3E06_IgG1 | This study | N/A |
| 011-10069 5C01_IgG1 | This study | N/A |
| 011-10069 5D01_IgG1 | This study | N/A |
| 011-10069 5G01_IgG1 | This study | N/A |
| 011-10069 5G04_IgG1 | This study | N/A |
| 013-10078 3G01_IgG1 | This study | N/A |
| 017-10116 5B03_IgG1 | This study | N/A |
| 017-10116 5D02_IgG1 | This study | N/A |
| 017-10116 5G02_IgG1 | This study | N/A |
| 019-10117 1B02_IgG1 | This study | N/A |
| 019-10117 3A06_IgG1 | This study | N/A |
| 019-10117 3C06_IgG1 | This study | N/A |
| 019-10117 3F01_IgG1 | This study | N/A |
| 019-10117 3F02_IgG1 | This study | N/A |
| 024-10128 3C04_IgG1 | This study | N/A |
| 024-10128 3F04_IgG1 | This study | N/A |
| 028-10134 4F03_IgG1 | This study | N/A |
| 030-121509 3B01_IgG1 | This study | N/A |
| 034-10040 4C01_IgG1 | This study | N/A |
| 034-10040 4E01_IgG1 | This study | N/A |
| 034-10040 4F02_IgG1 | This study | N/A |
| 034-100809 3E01_IgG1 | This study | N/A |
| 037-10036 5A01_IgG1 | This study | N/A |
| 041-10047 1C04_IgG1 | This study | N/A |
| 042-10065 2B03_IgG1 | This study | N/A |
| Software and Algorithms | ||
| GraphPad Prism 7.0 | GraphPad Software, Inc. | https://graphpad.com |
| PyMOL | Schrödinger | https://pymol.org |
| Other | ||
| Turkey whole blood in ACD | Lampire Biological Laboratories | 7209401 |
| Octet Red Biolayer instrument | ForteBio | N/A |
| Streptavidin (SA) Biosensors | ForteBio | #18-5019 |
| Immunospot ELISpot Imager | Cellular Technology Limited | N/A |