Premier
Immunization through Library Construction
Time: ~11-16 weeks
8-12 weeks
Phase I: Immunization
- 1 Llama
- 1 Pre-immune serum collection
- 4 Immunizations
- 3 Test serum collections
- ELISAs on each bleed
8-12 weeks
Phase II: PBMC Isolation
- PBMC isolation from week 8 and 12 bleeds
1 week
Phase III: cDNA Library Construction
- sdAb library as glycerol stock of phage plasmid-containing bacteria
Deliverables
- sdAb library as glycerol stock of phage plasmid-containing bacteria (optional)
Pro
Immunization through Final Clones
Time: 19-28 weeks
8-12 weeks
Phase I: Immunization
- 1 Llama
- 1 Pre-immune serum collection
- 4 Immunizations
- 3 Test serum collections
- ELISAs on each bleed
1 week
Phase II: PBMC Isolation
- PBMC isolation from serum collected at weeks 8 and 12
2-3 weeks
Phase III: cDNA Library Construction
- sdAb library construction from Phase II PBMCs
2-3 weeks
Phase IV: Library Panning and Screening
- 2 rounds of panning
- ELISA screening
4-6 weeks
Phase V: Single Domain Clone Selection and Sequencing
- Selection and sequencing of binders
- Small-scale antibody production
Deliverables
- Amino acid sequences of binders
- Purified antibody from up to 6 final clones
- Confirmed DNA sequences and glycerol stocks of final clones