Hybridoma Antibody Gene Sequencing
Hybridoma cells are prone to contamination, gene loss, poor cell status and even death during preservation, which will affect the quality of antibodies produced. The antibody gene in hybridoma cells was extracted and sequenced to obtain the corresponding nucleotide sequence, so as to facilitate the stable expression and production of exogenous antibodies in the later stage.
DetaiBio has the full-length mRNA sequencing platform and Failsafe ® Pseudogene Elimination technology, which can provide rapid and reliable hybridoma antibody gene sequencing services. Based on RACE (Rapid Amplification of cDNA Ends) technology, this service can accurately sequence the variable region or full length of the antibody. The sequencing range covers mice, rats, rabbits, goats and other species. At the same time, DetaiBio also has a mature antibody expression platform, which can express, purify and verify the obtained antibody genes.
Service Features
Extreme experience: PCR results can be obtained in 8 hours, and antibody subtypes can be obtained
Less cell demand: only 1~5 cells are needed, and pore plate samples can be received
Independent research and development: efficient enzymes and primers ensure 100% success rate when delivered in 5 days
Advanced technology: it can measure all subtypes of IgM and IgG of mice, rats, rabbits, goats and other species, and sequencing has no throughput limit
High accuracy: self built Failsafe® Pseudogene Elimination technology, which can be excluded ϰ Light chain pseudogene
One stop service: DetaiBio has rich experience and can provide expression verification services
Procedure
Workflow
| Stage | Service | Timeline | Deliverables |
|---|
| StageⅠ 1st Chain cDNA Amplification | - Cell lysis
- 1st chain cDNA amplification
| 5 days | - Detailed report
- Heavy&Light chain sequencing report
- Plasmid containing antibody fragment (optional)
- At least 0.5mg antibody SDS purity ≥ 90% (optional)
|
| Stage Ⅱ Antibody Gene Amplification | - Gene pre amplification
- Segmented gene specific amplification
|
| Stage Ⅲ Sequencing | - Cloned to vector
- Sanger sequencing
- Bioinformatic analysis
|
| Stage Ⅳ Transient Transfection Expression & Purification (optional) | - Transfection level plasmid preparation
- Transient transfection expression
- Cell sample collection
- Affinity Purification
- QA/QC
| 7 days |
| Stage Ⅴ Subcloning (optional) | - Subcloning with the plasmid of stage III as the template
- The expression plasmids of light and heavy chains were constructed respectively
| 4 days |
Note：Customers need to provide hybridoma cells in good condition
Case Study
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