August 27, 2013 -- Creative BioLabs has established a solid platform for DNA sequencing of both IgG and IgM types of monoclonal antibodies produced by mouse and rat hybridoma cell lines. The sequences of a monoclonal antibody are important for patent protection and therapeutic approval. Starting from a hybridoma cell line, Creative Biolabs offers the following services:
1. Validation of the hybridoma cell line in terms of antibody production, antibody isotyping and antigen-binding specificity;
2. RNA extraction and reverse transcription;
3. Full length IgG or Fab sequencing by PCR amplification and subcloning of the variable domains.
Features:
The ability to clone right light chain The cloned cDNA fragments are derived from the antigen-binding antibody Express the sequenced scFv/Fab validate its authenticity in an ELISA assay Express the sequenced VL and VH in full IgG format to ensure the uniform binding specificity and affinity Keep the original FR1 sequence Guarantee successful services otherwise refund payments It is noted that hybridoma cells may contain pseudo genes and mRNAs encoding non-functional antibody chains, which might be accidentally amplified by the primer pairs intended to clone the target antibody sequences. Therefore, in a total RNA [as well as cDNA] pool of the hybridoma cells, there are other antibody-like sequences [e.g. other IgG] that can be amplified along with the sequences of the target antibody. In fact, PCR amplifications intended to clone coding sequences [cDNAs] of the mouse/rat monoclonal antibody will inevitably amplify some other antibody-like sequences. This is the key challenge in sequencing the cDNAs of a particular monoclonal antibody. In extreme cases, a small phage display scFv/Fab library is constructed for each monoclonal antibody, and then the right scFv/Fab clones are selected against the original antigen.
For antibodies intended for therapeutic use or sequencing the N-terminals of the heavy chain and the light chain using protein sequencing methods to confirm the cDNA sequences cloned from hybridoma cell lines, you can contact info@Creative-Biolabs.Com or call 1-631-871-5806.
For details, information can be reached at http://www.creative-biolabs.com/antibody-sequencing-service.html
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