Antibody Sequencing Services
Fusion Biolabs offer fast and reliable antibody sequencing services for monoclonal antibodies. Our unique technology allows us to sequence your antibodies from any antibody producing cell line (hybridomas, peripheral blood lymphocytes, B-cells, spleen cells or bone marrow), and from a variety of species (mouse, rat, hamster, rabbit, chicken, camelids, human and other primates). The service can be tailored to specific parameters, from variable region or domain to full-length antibody sequencing.
Why sequence your monoclonal antibody (mAb)?
- Patent Application – If you plan to obtain a patent for your mAb, antibody sequencing will be necessary in order to distinguish your novel antibody from others.
- Antibody Recombination – To determine the sequence of a mAb, the cDNA of your antibody can be isolated from a hybridoma cell line and its subcloning and expression in mammalian cells.
- Antibody Humanization – Antibody sequence data is essential for any antibody engineering or humanization development.
What We Offer
Variable Region Cloning and Sequencing
- Total RNA extraction from hybridoma cell pellets or other sources (spleen, B-cells, etc.)
- Reverse transcription
- PCR using degenerate primers for variable domains
- Cloning of variable heavy chain sequence
- Cloning of variable light chain sequence
Leader Sequence & Variable Domain Cloning and Sequencing
- Known sequences from Variable Domains Cloning and Sequencing Services
- 5’RACE amplification of heavy and light chain from antibody constant domains CH1 and CL
- Cloning of variable heavy chain sequence
- Cloning of variable light chain sequence
Full-Length Antibody Cloning and Sequencing
- Known sequences from Leader Sequence & Variable Domain Cloning and Sequencing Services
- 3’RACE amplification of the heavy and light chain from antibody constant domains CH1 and CL
- Cloning of variable heavy chain sequence
- Cloning of variable light chain sequence
Key Features
- 100% success rate
- Quick turnaround
- Fully annotated antibody sequences at DNA level
- Fully annotated antibody sequences at amino acid level, which include complementarity-determining regions (CDRs) and framework annotation
Process Overview
Starting Materials
1) Pellet of snap frozen hybridoma or B-cells (1×105-1×107 cells) and other sources expressing your antibody.
2) Additional information desired: isotype of starting antibody (Download and fill Oak Biosciences Antibody Sequencing Order Form)
Phase 1: Total RNA Extraction from Hybridoma Cell Pellet
Total RNA will be extracted and purified from hybridoma cell pellet. The quality of the total RNA will be analyzed using Agilent Bioanalyzer 2100.
Phase 2: Reverse Transcription
Total RNA will be transcribed into cDNA using either an oligonucleotide (dT) or a gene-specific anti-sense primer.
Phase 3: PCR or RACE Amplification of Heavy and Light Chains
Proprietary primers are used to amplify the variable domains from cDNA. For RACE, our proprietary adaptor is added to the end of cDNA allowing heavy and light chains to be amplified with our proprietary adaptor and a specific primer. PCR products will include the sequence of signal peptide, variable domains and constant domains. It could also be used to determine the isotype of the antibody.
Phase 4: Cloning Into a Standard Sequencing Vector
The PCR products will be gel purified to clone into a sequencing vector for sequencing.
Phase 5: Sequencing Analysis
A minimum of 12 independent clones for each chain will be sequenced.
Phase 6: Final Report
A comprehensive report consisting of fully annotated antibody DNA, protein sequence, and CDRs will be provided.
Deliverables
1) CDR and framework annotated sequences of both heavy chain and light chain
2) Sequencing plasmids with inserted regions (VH, VL, CH and CL)
Ordering Information
Antibody Sequencing and Cloning Services for Single B cells and Hybridoma
| Catalog | Sequencing Regions and Timeline | Sources | Unit | Price |
|---|---|---|---|---|
| 80101 | Variable Domains (VH and VL) Sequencing for both Heavy Chain and Light Chain; 5 days | Single B Cell or Hybridoma or Spleen from Human/ Murine / Rat / Rabbit/ Hamster/ Chicken | 1 Project | Contact Us |
| 80102 | Leader region + Variable Domains Sequencing for both Heavy chain and Light Chain; 10 days | Single B Cell or Hybridoma or Spleen from Human/ Murine / Rat / Rabbit/ Hamster/ Chicken | 1 Project | Contact Us |
| 80103 | Full length Antibody Sequencing (Leader sequences + Variable regions + Constant regions CH and CL) for both Heavy chain and Light Chain; 12 days | Single B Cell or Hybridoma or Spleen from Human/ Murine / Rat / Rabbit/ Hamster/ Chicken | 1 Project | Contact Us |
| 80104 | Antibody Repertoire Deep Sequencing (NGS); 14 days | Single B Cell or Hybridoma or Spleen from Human/ Murine / Rat / Rabbit/ Hamster/ Chicken | 1 Project | Contact Us |
For complex project, please fill the form below for a customized quotation or proposal.