Antibody Production
Proteos provides 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 10 grams of antibody in a single production run, buffer exchange by dialysis, endotoxin testing, analytical SEC, and SDS-PAGE (reducing and non-reducing).
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 also offer cell line expansion, cryopreservation, and banking of client-provided cell lines. Master banks will be maintained exclusively for the client’s transient or stable protein production projects and will be destroyed or returned at their request.
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 batch size. 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 antibody production starting from construct design.
Full production services:
- Full suite of molecular biology services to accompany any of our expression platforms, including construct design, fusion partner and affinity tag selection, gene synthesis with codon optimization, subcloning, endotoxin-free plasmid DNA isolation, and sequence verification.
- Expression verification and optimization at culture scales less than 100 mL; ideal for expression testing multiple antibodies in parallel.
- Medium-throughput production of sets of 8 antibodies in parallel at a 30 mL culture scale. Up to 48 antibodies may be produced in parallel.
- Pilot scale production and yield determination at culture scales ranging from 1 L to 4 L; ideal for process verification through purification prior to large-scale production.
- Preparative scale production of recombinant full length IgG, antibody fragments, or antibody fusion proteins in HEK or CHO cells in shake flask culture volumes up to a 60 L batch size.
- Antibody expression services can be combined with Proteos’ purification services for a fully-customized protein production process.
Additional antibody 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 BSL-1 biological fluids
- Antibody isotype determination
- Antibody labeling (HRP, biotin, FITC, etc.)
- Antigen production