Key Indicators of Human Fab Naïve Library
Source | ~1000 healthy donors | Material | Human PBMC |
Library | Fab phage library | Total Size (cfu) | 1.0x1011 (in expansion) |
Insertion rate | >95% | In-frame rate | >85% |
Diversity rate | >95% | CDR3 diversity | Normal distribution |
Tag | His & c-Myc | Affinity with SPR | 1E-8~1E-10 |
Up to 2 panning strategies to maximize the success rate
Two step validation: monoclonal phage ELISA and soluble ELISA
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ProBio selected antibody molecules targeting CLDN18.2 from our human Fab naïve library using a combination of ELISA and FACS screening, reverse screening and epitope identification against homologous family member Claudin 18.1, and ADCC/CDC assays. As a result, 11 superior antibody molecules were identified with the following characteristics:
Criteria | Lead | BMK-IMAB362 |
---|---|---|
ELISA EC50 | 0.07 | 1.53 |
FACS EC50 with Claudin18.2 | 0.71 | 3.97 |
FACS EC50 with Claudin18.1 | NA | NA |
Affinity with SPR | 2.70E-09 | 3.93E-08 |
ADCC | 0.38 | 0.76 |
CDC | 1.12 | 6.96 |
Epitope binning | / | same |
In vivo assay | ongoing |
Screening the antibody leads targeting PD-L1 with ProBio‘s human Fab naïve library. The leads were narrow down to 4 dominant antibody molecules with ELISA&FACS EC50, species cross-reactivity screening, blocking screening with the natural receptor PD1, and epitope binning:
Criteria | Lead(best) | BMK1-Avelumab | BMK2-Tencentriq |
---|---|---|---|
ELISA EC50 | 0.2 | 0.7 | / |
FACS EC50 | 0.01 | 0.07 | / |
Affinity with SPR | 3.05E-10 | 8.34E-10 | / |
Cross-reactivity with Mouse | 0.02 | 0.04 | / |
Cross-reactivity with Cyno | 0.11 | 0.13 | / |
Epitope binning | / | Similar and different | Similar and different |
PD1/PD-L1 blockade assay | 0.16 | 1.41 | 0.38 |
In vivo assay | ongoing |