Antibody Production
Proteos offers production of full-length IgG antibody, F(ab) and (Fab’)2 antibody fragments, and antibody fusion proteins. Our antibody production capabilities include construct design, transient mammalian cell expression, utilizing in-house or customer provided expression vectors, and affinity purification. Additional purification services offered include size exclusion chromatography of up to 5 grams of antibody in a single production run, buffer exchange by dialysis, endotoxin testing, analytical SEC, SDS-PAGE (reducing and non-reducing), and sterility testing.
Our industry leading transient expression platform utilizes fully licensed HEK (human embryonic kidney) and CHO (Chinese hamster ovary) cells lines expressing the Epstein-Barr virus (EBV) nuclear antigen 1, with proprietary expression vectors containing the EBV origin of replication (OriP) to allow episomal plasmid amplification and high level recombinant protein expression.
We utilize optimized and fully validated mammalian cell expression protocols for consistent, reproducible protein expression in serum-free shake flask culture volumes ranging from 30 mL to 60 L per batch. Only mycoplasma-free, high viability, low passage cultures are used for transfection and are monitored using a Beckman Coulter Vi-cell XR for culture density, viability, and average cell diameter. The Vi-cell XR uses the trypan blue dye exclusion method and averages 50 measurements to compute the parameters for each sample. This allows us to monitor cultures with a high degree of accuracy which is imperative for the quality and reproducibility of protein production projects. All cell culture data is included in a customized technical report delivered with each project.
Typical productions times are 6-8 weeks for construct design, expression, and purification.
Full production services:
- Proteos offers a full suite of molecular biology services for antibody production which includes: construct design, codon-optimized synthesis, subcloning, endotoxin-free plasmid isolation, and sequence verification.
- Small-scale production optimization at scales less than 100 mL.
- Medium-throughput production of up to 48 antibodies in parallel at the 30 mL culture scale
- Analytical production and yield determination at scales up to 4 L; ideal for process verification through purification prior to large-scale production.
- Production of recombinant full length IgG, antibody fragments, or antibody fusion proteins in HEK293-6E or CHO-3E7 cells up to 60 L per batch in shake flasks.
Additional services:
- Enzymatic processing of full length IgG for production of F(ab) or F(ab’)2 antibody fragments
- IgG purification from sera, ascites, or other biological fluids
- Antibody isotype determination
- Antibody labeling (HRP, biotin, FITC, etc.)
- Antigen production