Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Pseudomonas aeruginosa) | PA103 | ATCC | Cat#:29260 | sputum isolate |
| Cell line (human) | HEK-293T (epithelial-like, kidney) | ATCC | Cat#:CRL-3216; RRID:CVCL_0063 | Cell line transfected for antibody production |
| Transfected construct (human) | Antibody expression plasmids (plasmid) | Addgene | RRID:Addgene_183702 (kappa light chain);RRID:Addgene_183703 (lambda light chain); RRID :Addgene_80795 (IgG1 heavy chain) |
Deposited in Addgene by Dr. Hedda Wardemann |
| Biological sample (human) | PBMC | Seattle Children’s Cystic Fibrosis Clinic | Collected from individual donors under IRB approval SCH#10325 | |
| Antibody | HRP-conjugated anti-human IgG; IgM (goat polyclonal) | SouthernBiotech | Cat#:2040–05 (IgG); Cat#: 2020–05 (IgM) | ELISA IgG (1:3000); IgM (1:1500) |
| Antibody | HRP-conjugated anti-human IgA (goat polyclonal) | Thermo Fisher Scientific | Cat#:A18781 | ELISA (1: 1:1500) |
| Recombinant DNA reagent (Pseudomonas aeruginosa) | pET16b PcrV (plasmid) | Sawa et al., 1999 | Plasmid encoding histidine-tagged PcrV; gift from Dr. Timothy Yahr | |
| Peptide, recombinant protein | Streptavidin conjugated to fluorophore | Agilent | Cat#:PJ27S (SA-APC); Cat#:PJRS25 (SA-PE) | For labeling of protein tetramer |
| Commercial assay or kit | In-Fusion HD Cloning | Takara Bio | Cat#:639650 | |
| Commercial assay or kit | EZ-Link Sulfo-NHS-LC Biotinylation Kit | Thermo Fisher | Cat#:21435 | |
| Commercial assay or kit | SMART-seq v4 | Takara Bio | Cat#:634770 | cDNA synthesis |
| Chemical compound, drug | gremubamab | Invitrogen/Thermo Fisher Scientific | Cat#:MA5-42275 | |
| Software, algorithm | IgBlast | Ye et al., 2013 | ||
| Software, algorithm | IGMT/HighV-Quest | Alamyar et al., 2012 | ||
| Software, algorithm | Prism | GraphPad | Version 9.5 | |
| Software, algorithm | TRUST4 | Song et al., 2021 | ||
| Other | Amicon Ultracentrifugal Filter | Sigma Aldrich | Cat#:UFC9050 | For concentration and buffer exchange of purified mAbs |
| Other | HiTrap Protein G HP purification column | Cytiva/Thermo Fisher Scientific | Cat#:45-000-053 | For purification of IgG from concentrated transfectant supernatant |
| Other | POROS CaptureSelect IgM-XL Affinity Matrix | Thermo Fisher Scientific | Cat#:2812892005 | For purification of IgM from concentrated transfectant supernatant |