Phase 1: Project design
Analyze the antibody protein sequence and determine the functional regions for gene expression. Design antibody expression formats (VHH, scFv, HC-LC, etc.) to meet the client’s requirements.
Lead Time: 1-2 days
Phase 2: Gene synthesis and cloning
Chemically synthesize antibody genes and optimize the gene codons for a mammalian expression system to improve gene expression. Alternatively, use the customer-provided plasmid as a gene template and clone the target genes into our mammalian cell overexpression vector. The recombinant vectors will be confirmed by DNA sequencing.
Lead Time: Usually 2-4 weeks
Deliverable: DNA sequencing chromatography data.
Phase 3: Small-scale gene expression
The plasmid DNA is transfected into a small-scale culture of CHO suspension cells, typically 2 ml. We examine the secreted antibodies in the cell culture supernatant via SDS-PAGE and western blotting to evaluate gene expression levels. The sample’s raw activity is confirmed by us or the customer via ELISA or other immunological methods. If the recombinant antibodies exhibit high expression levels and the expected functional activity, the project will proceed to Phase 4.
Lead Time: Usually 1-2 weeks
Deliverable: SDS-PAGE and western blotting results.
Phase 4: Large-scale gene expression and purification
Based on the desired amount of antibody, scale up CHO cell transient transfection, starting from 0.5 liters to 10 liters, or more. Extract large amounts of plasmid DNA and transfect CHO cells on a greater scale. Alternatively, we can create stably expressing cell lines to produce bulk recombinant proteins for your needs (additional lead time may apply).
Lead Time: Usually 2-4 weeks
Phase 5: Recombinant antibody affinity purification
The cell culture supernatants of the transfected cells are collected for antibody affinity purification. The purified antibody can be aliquoted, lyophilized, and labeled upon request (additional lead time may apply).
Lead Time: Usually 1-2 weeks
Deliverable: All antibodies purified in this phase and, if applicable, the stably expressing cell line.