Hybridoma Sequencing Service
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Workflow
Extraction of Hybridoma mRNA
Hybridoma cells are harvested from culture, and RNA is extracted using a kit. The focus is on isolating mRNA, particularly the antibody-related transcripts. The RNA quality is checked to ensure it’s suitable for sequencing.
High-Throughput Sequencing
The extracted mRNA is converted into cDNA. A sequencing library is prepared, and the cDNA is sequenced using high-throughput NGS platforms. The quality of the sequencing data is then checked to ensure accuracy.
Transcript Assembly and Refinement
The raw sequencing data is assembled into transcript sequences without a reference genome. The assembly is optimized by removing low-quality sequences and duplicates. The final set of transcripts is validated for accuracy.
Search for lgH, lgL, and lgK Sequence
The assembled data is searched for heavy chain (lgH), light chain lambda (lgL), and kappa (lgK) sequences. The variable and constant regions are annotated, and the sequences are confirmed to be functional by comparing with antibody databases.
Service Description
Hybridoma cells are highly sensitive to contamination and suboptimal conditions, which can lead to the loss of antibody genes. To prevent this, timely extraction and sequencing of these genes are essential. By obtaining precise gene sequences, we can utilize recombinant antibody expression for consistent production, ensuring the long-term preservation of these valuable antibodies.
At Bon Opus BioScience, our hybridoma antibody sequencing service efficiently captures monoclonal antibody (mAb) genes, followed by optimized codon usage and gene synthesis strategies. This allows us to express and purify antibodies with high quality and reliability for our clients.
Our hybridoma services include:
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Validation of hybridoma cell lines for antibody production, antibody isotyping, and antigen-binding specificity
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RNA extraction and reverse transcription
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scFv sequencing through PCR amplification and subcloning of the variable domains
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Full antibody sequencing, including subcloning both variable and constant domains
Step
1
2
3
Working Day
7
6
1
Service Type
Gene synthesis (variable region, VH/VL), subcloning into respective constant vectors
Transfection-grade plasmid prep, HTP transfection of 293/CHO cells
H-T purification (magnetic Protein A beads)
Altum Mab™ High Throughput Package ( Start from $385 ) * minimum 10 pairs to start *
Deliverables:
200μg mAb option (>95% purity by SDS-PAGE), plasmid
1 mg mAb option (>95% purity by SEC-HPLC, endotoxin level <1EU/mg), plasmid
Total: 14 Days*
Step
1
2
Service Type
Gene synthesis (variable region, VH/VL), subcloning into respective constant vectors
Transfection-grade plasmid prep
Working Day
7
3-5
Altum Gene™ High Throughput package * minimum 50 constructs to start *
Deliverables:
Up to 100ug transfection-grade DNA, Sequencing report
Total: 10-12 Days*
*Shipping time is not included
Here at Bon Opus Biosciences, we treat each request and deliverable with the highest level of integrity. Our experienced and devoted team of field scientists and researchers pride themselves on their ability to offer the most up-to-date biotech and sciences, and that now includes high throughput antibody production services. We understand the need to gain awareness and insight into scientifically backed and proven approaches, and our in-house team is proud to offer a custom range of services to each of our partnerships. We have developed, tested, and mastered the high throughput antibody production process, which provides us with the opportunity to remain collaborative, transparent, and affordable. We are happy to offer comprehensive antibody expression packages, in addition to our custom services, per partnership demands. A member of our highly trained and educated team will work closely with you through each step to ensure the package is right for your needs and the customization process is seamless. To learn more about how Bon Opus Biosciences can serve you in collaboration and recombinant proteins, fill out the online contact form, and a member of our team will be in communication shortly.
WHAT PEOPLE SAY
Dr. Thomas B. Kepler, Professor
Department of Microbiology
Boston University School of Medicine
The antibodies you made worked out very well. Thank you for your excellent service! - June, 2022