Monoclonal Sequencing Service  - click here for brochure

Fusion Antibodies offers a fast and professional antibody sequencing service for your valuable monoclonal antibodies.  We can sequence from any antibody producing cell line with a service that can be tailored to your requirements.

Whatever your application the ISO 9001:2000 certified antibody sequencing service is one that you can depend on.
 
Quality Control              Third party verification of your target antibody cell line gives you 
                                          added security in your processes

Patent Applications     Sequence information can support and strengthen your patent

Full Traceability             Ensure batch to batch consistency throughout your research,
                                          development and manufacturing programs

Drug Development       Sequence data can assist in the selection of target cell line and is vital
                                          for any downstream antibody engineering.

Each antibody cell line project comes complete with a full comprehensive report of the alignments.

Antibody service Levels

Standard Sequencing Service - Variable Domains - 3 weeks

5'RACE Sequencing Service - Variable Domains & Leader Sequence - 4 weeks

Full Sequencing Service - Variable Domains & Constant Domains - 5 weeks


Please enquire for more details on this service and pricing with our technical services team at  technicalservices@fusionantibodies.com or call us on  +44(0) 28 9043 2800

 

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 Technical Summary

Stage 1: mRNA extraction from the hybridoma cell pellet
mRNA will be extracted and purified from the hybridoma cell pellet using standard protocols.

Stage 2: Reverse transcription
mRNA will be transcribed into cDNA using either an oligo dT anti-sense primer or a gene-specific (murine or human IgG1 CH and kappa CL) anti-sense primer. The initial cDNA production will most probably be with the oligo dT primer. Specific murine and human constant domain primers could be used to PCR after cDNA production to determine the isotype of the antibody.

Stage 3: PCR or 5’ RACE amplification of heavy and light chains
Degenerate VH and VL primers are used to amplify the variable domains from the cDNA.
For 5’RACE, a homopolymeric dCTP tail is added to the 3’ end of the cDNA. The heavy and light chains can now be amplified with an oligo dG sense primer and a gene specific (CH/CL) anti-sense primer. PCR products will include the sequence of the signal peptide, variable domains and constant domains up to the anti-sense primer.

Stage 4: Cloning into a standard sequencing vector
The PCR products will be gel purified to remove small fragments, and cloned into a blunt or TA vector for sequencing.

Stage 5: Sequencing analysis
The specified number of clones to be sequenced can be determined by the client. As standard we will sequence a minimum of 5 independent clones for each chain.

Stage 6: Final Report
A report is put together to detail the work performed which encompasses all the blots and sequence alignments of the heavy and light chains and is e-mailed to the client.