Full Coverage Mammalian Expression System for Affinity Maturation
FCMES-AM™ (Full Coverage Mammalian Expression System for Affinity Maturation) is Biointron's proprietary platform for affnity maturation. For each amino acid in CDR region, it will be mutated to the other 17 amino acids (except Cysteine and Methionine) with equal ratio. The clones will be expressed in mammalian system and ELISA binding assay will be conducted as initial test, followed by SPR or FACS for further affinity measurement.
| Milestone | Service Description | Deliverables |
|---|---|---|
Sequence Design | No-bias Site Saturation Mutagenesis in CDR Region (all 17 amino acid variants for each amino acid in CDR region ) | / |
Hot Spots Identification | ▸High Throughput Expression in Mammalian System (FCMES-AM™) ▸ ELISA Detection | ELISA Results |
Hot Spots Combination | ▸ High Throughput Expression in Mammalian System (FCMES-AM™) ▸ ELISA Detection | ELISA Results |
Antibody Expression & Affinity Detection | ▸Antibody Expression & Affinity Purification ▸Affinity Detection by Biacore Full KD or FACS | ▸ Purified Antibody ▸ Plasmid Sequencing Results ▸ Affinity Detection Results |
Koff of the VHH improves and the affinity increases 6-fold, 23-fold and 19-fold separately.
Affinity increases 120-fold, 48-fold and 46-fold separately.
