GMAb has responded to customer needs and optimized our antibody sequencing to provide highly accurate hybridoma cell line sequencing with the fastest turnaround time on the market. Our optimized approach, processed in our newly expanded laboratory, makes sequences available in an average of 3 business days.
Sequence your antibody to safeguard from the possibility of losing the hybridoma cell line. This critical piece of information is required for patent applications. Once your sequence is identified, we can express the sequence recombinantly to validate binding and create the ideal format for your application.
Our Approach
All antibodies have the same basic protein structure but exhibit sequence variability in the regions that contact the antigen. Our variable domain sequencing identifies these VH and VL sequences in the heavy and light chain of the immunoglobulins from the mRNA encoding the antibody and expressed in the hybridoma cell line.
The antibody sequencing services at Green Mountain Antibodies can be completed in as fast as 3 days from the receipt of your cell pellet. See our sample report.
Why Green Mountain Antibodies?
Capture the entire variable domain: By utilizing rapid amplification of cDNA ends (RACE), we capture the entire variable domain plus the leader sequence and 5’ UTR, ensuring that we report every residue in the variable domain that makes YOUR antibody unique.
Optimized sequencing approach: Nanopore sequencing combines the best features of other sequencing technologies – the many reads of NGS (allowing us to see the complexity of the hybridoma) with the speed of Sanger sequencing (overnight results). Plus, length is no longer a limiting factor, so we can easily cover as much of the sequence as needed. Improvements in ONT has made the sequence highly accurate (>99.99% with >15 reads). We have extensively compared with other approaches and see 100% sequence agreement.
We know more: Every sequence is analyzed by an experienced molecular biologist to verify functional antibody sequences and ensure no process contamination. Sequence your entire project with us to get in depth phylogenetic comparison and summary of sequence liabilities. If your hybridoma is identified as complex, we can express your sequences recombinantly, assay them here, and tell you exactly which heavy and light chains affect binding (plus some insights on developability).
Speed: Biotechnology is a fast-paced field, and your time is important. That’s why our antibody sequencing processes are built for speed, while still providing the highest level of accuracy. Our average turnaround time for standard VH/VL sequencing is 3 business days from pellet to report.
Service Details
We accept cell pellets or frozen vials of hybridoma cells or clonal B cells. Ask us for detailed instructions on alternative methods to submit your sample. GMAb can also culture your cells for the sequencing process, so you can avoid sample preparation time at your facility.
Our experts can design schemes for all your sequencing needs, including antibodies from any species (mouse, rat, human, rabbit, hamster, camel, sheep, etc.) and any constant region (IgG1, IgG2a, IgG2b, IgG3, IgG4, IgM, IgE, IgA, IgK, IgL, etc.) Let us know what you need and we’ll design a solution.
Applications
- Hybridoma identification
- Antibody engineering
- Expression in alternate platforms
- Antibody humanization
- Confirm diversity
- Antibody binning
- Antibody immortalization and preservation
