• Consultation in peptide antigen design
• Peptide antigens with post-translational modifications can be made
• Flexible protocol and schedule based on customer’s needs
• Immunization time frame of four to eight weeks
• Up-to-date report and customer validation following each project step
• Accessible technical support with expertise in antibody production
• Resources and technical proficiency to work with customer on unconventional projects
Our routine production of monoclonal antibodies is comprised of three phases (Phase I – III). Customers only pay for each phase that is successfully completed. Phase IV, large-scale production of monoclonal antibodies, is optional and available as a stand-alone purchase. Details of each phase are shown below.
Phase I: Antigen Preparation and Immunization (~8-9 weeks)
• Design and production of synthetic peptide antigens with or without modifications
• Site-directed conjugation of peptide antigens to carrier proteins (e.g., KLH and BSA) for flexibility in antigen presentation
• Preparation of peptide and protein antigens with adjuvants for immunization
• Initial and boost immunizations of at least four animals
• Serum separation, ELISA testing and customer consultation before proceeding to Phase II Phase II: Fusion and Selection of Parental Clones (~3 weeks)
• Splenocytes are harvested for cell fusion from mice with the highest titer
• Clones of fused cells are expanded and screened by ELISA.
• Supernatants, from up to ten ELISA-positive parental clones, are shipped to customer for further antibody testing in immunoassays
• The best parental clones (up to three) are chosen for Phase III after customer consultation
Phase III: Subcloning and selection of Single Clones (~3 weeks)
• Parental clones are sub-cloned by limiting dilution to isolate single clones
• Isotype determination of single clones – IgG subtype guaranteed single clones are expanded and screened by ELISA
• Single clones with high specificity (e.g., phospho vs. non-phospho) and antibody titer will be expanded and cryopreserved
• Three hybridomas, and their supernatants, will be shipped to customer
Phase IV: Large-Scale Production of Monoclonal Antibodies (optional) (~3-6weeks)
• In vivo ascites or in vitro Bioreactor antibody production is available
• Purification of monoclonal antibodies by affinity chromatography using Protein G or with the immunizing peptide antigen
• ELISA and SDS-PAGE are done to validate the purity of the antibody
• AnaSpec offers a range of production scales with flexible cost adjustment
In addition to our custom services, AnaSpec, EGT Group offers popular catalog monoclonal antibodies such as the anti-amyloid (1-40) and (1-42) antibodies.
For more information, please visit http://www.anaspec.com/products/promotions.asp?col=1&row=1
AnaSpec, EGT Group
AnaSpec, Eurogentec Group is a leading provider of integrated proteomics and genomics solutionsTM for worldwide life science research. AnaSpec offers expertise in peptides, detection reagents, antibodies, assay kits, amino acids, oligonucleotides, and qPCRs. AnaSpec carries a broad product line of biochemicals and reagents for basic research, high-throughput screening and drug discovery. AnaSpec also provides premier custom services including peptide synthesis, antibody production, assay development, fluorescent dyes, and oligonucleotide synthesis. AnaSpec is certified to ISO 9001:2008.
For more information visit www.anaspec.com
