organism: Vicugna pacos
attributes.tag: isolate
attributes.value: 'Prince'
attributes.tag: collection date
attributes.value: 2022-10-17
attributes.tag: geo_loc_name
attributes.value: USA: New Haven
attributes.tag: isolation source
attributes.value: phage display library
attributes.tag: synthetic
attributes.value: True
attributes.tag: sex
attributes.value: male
attributes.tag: age
attributes.value: not collected
attributes.tag: dev_stage
attributes.value: adult
attributes.tag: age_event
attributes.value: sampling
attributes.tag: population
attributes.value: not applicable
attributes.tag: ethnicity
attributes.value: not applicable
attributes.tag: race
attributes.value: not applicable
attributes.tag: strain
attributes.value: not applicable
attributes.tag: study_group_description
attributes.value: extended high-throughput cell-based phage display panning
attributes.tag: disease
attributes.value: not applicable
attributes.tag: disease_length
attributes.value: not applicable
attributes.tag: disease_stage
attributes.value: not applicable
attributes.tag: prior_therapies
attributes.value: not applicable
attributes.tag: treatment
attributes.value: not applicable
attributes.tag: medical_history
attributes.value: not applicable
attributes.tag: tissue
attributes.value: peripheral blood lymphocytes
attributes.tag: anatomic_site
attributes.value: peripheral blood
attributes.tag: health_state
attributes.value: not applicable
attributes.tag: collection_time_point_reference
attributes.value: panning_start
attributes.tag: collection_time_point_relative
attributes.value: 348
attributes.tag: collection_time_point_relative_unit
attributes.value: id: UO:0000033, label: day
attributes.tag: biomaterial_provider
attributes.value: Barbara Kazmierczak, Yale University, Ste S140, 300 Cedar Street, New Haven, CT 06519
attributes.tag: tissue_processing
attributes.value: phage elution by hydrochloric acid/triethylamine elution
attributes.tag: cell_type
attributes.value: phage display
attributes.tag: cell_phenotype
attributes.value: not applicable
attributes.tag: single_cell
attributes.value: False
attributes.tag: cell_number
attributes.value: not collected
attributes.tag: cells_per_reaction
attributes.value: not collected
attributes.tag: cell_storage
attributes.value: Y
attributes.tag: cell_quality
attributes.value: not collected
attributes.tag: cell_isolation
attributes.value: not applicable
attributes.tag: target_antigen
attributes.value: MexEF-OprN
attributes.tag: phage_display_bait_type
attributes.value: bacterial cells
attributes.tag: selection_name
attributes.value: 027j.1.B4-.1
attributes.tag: counter_selection_cell_genotype
attributes.value: ∆{mexAB,oprM} +nfxB ∆{mexCD,oprJ} ∆mexJKL ∆mexXY ∆opmH362 ∆{mexEF,oprN}
attributes.tag: counter_selection_cell_background
attributes.value: PAO1 PA0397
attributes.tag: counter_selection_cell_strain_number
attributes.value: 1468
attributes.tag: counter_selection_cell_growth_condition
attributes.value: exponential phase
attributes.tag: counter_selection_cell_growth_protocol
attributes.value: a single colony was used to inoculate a culture of LB plus plus arabinose 0.2% w/v which was grown shaking at 37 °C overnight, then subcultured 1/25 into the same media and grown for 3 hours.
attributes.tag: selection_cell_genotype
attributes.value: +mexEF+oprN
attributes.tag: selection_cell_background
attributes.value: PAO1 PA0397
attributes.tag: selection_cell_strain_number
attributes.value: 1471
attributes.tag: selection_cell_growth_condition
attributes.value: exponential phase
attributes.tag: selection_cell_growth_protocol
attributes.value: a single colony was used to inoculate a culture of LB plus plus arabinose 0.2% w/v which was grown shaking at 37 °C overnight, then subcultured 1/25 into the same media and grown for 3 hours.
attributes.tag: technical_replicate
attributes.value: 1
attributes.tag: cell_processing_protocol
attributes.value: Library of phage-displayed VHHs were applied to counter-selection bacterial cells pre-washed with PBS+Ca+Mg and incubated 1 hour; cells collected by centrifugation 3750 rcf x 5 min and supernatant removed; counter-selection bacteria washed washed 5 times with PBS + Ca + Mg. Phage were eluted from counter-selection bacterial cells by addition of 0.1 N HCl x 5 min, neutralization with 1 M Tris-HCl pH 11; bacteria harvested and second elution performed with 0.1 M triethylamine (TEA) x 5 min and neutralization with Tris pH 6.8. Acid- and base- eluates combined. 10 µL of eluate used as template.
attributes.tag: immunogen
attributes.value: Four P. aeruginosa laboratory strains---PA14 (serotype O10), PAO1 (O5), PAK (O6) and PA103 (O11)---were grown to exponential phase in liquid LB and overnight on LB agar; cells were harvested, lysed by french press (>14,000 psi), and the lysate clarified by centrifugation. Membrane proteins were enriched by centrifugation through sucrose density gradients. Additionally, PA14 and PAO1 were grown as static biofilms in petri dishes in both LB and M9 minimal media at 30 ˚C for 36 hours, harvested by scraping, and lysed as above. The soluble proteins, enriched membrane proteins, and biofilm extracts were pooled to generate a mixed antigen preparation.
attributes.tag: phage_display_selection_round
attributes.value: 6
attributes.tag: phage_display_selection_input_or_output
attributes.value: output
attributes.tag: phage_display_selection_selection_or_counterselection
attributes.value: selection
attributes.tag: BioSampleModel
attributes.value: Model organism or animal