Antibody Sequencing
Monoclonal Antibody Sequencing
Poochon offers a rapid cDNA based de novo sequencing service for your valuable monoclonal antibodies. We can sequence from any antibody producing cell line with a service tailored to your requirements for quality control, patent applications, antibody engineering, and drug development. Our monoclonal antibody sequencing services are for sequencing of the variable domains of the heavy and light chain, the leader plus variable domains of the heavy and light chain, or the full length heavy and light chain from hybridomas and clonal B cells. Antibody types include IgG, IgM, IgA, and IgE antibody variable regions (VH and VL) from mouse, rat, hamster, rabbit, monkey, human, llama, alpaca, and camel. Plus, we offer protein amino acid sequence confirmation by LC-MS/MS.
+ A101: Antibody Sequencing of Variable Domains
Sequencing of Variable Regions, Plus Leader Sequences of Both Heavy and Light Chains, From mAb Producing Cell Lines
Price: $1800 per sample
Turnaround Time: 10 to 15 business days
Service Description:
• Extraction of total RNA from the cell pellet
• Reverse transcription and PCR amplification of heavy and light chains by 5′-RACE and 3′- RACE
• Specific murine and human constant domain primers are used to determine the isotype of the antibody
• Amplified products are purified and inserted into a sequencing vector
• Multiple independent colonies are sequenced for each chain to ensure accuracy (typically 8 colonies per chain)
Deliverables:
• Detailed project report with heavy chain variable region (VH) and light chain variable region (VL), plus leader sequences and alignments
• Plasmids with VH or VL, plus leader sequences
Suitable Sample Type:
•Frozen cells (1×107 cells) expressing target mAb
• EBV transformed B-cell line
• IgG expressing CHO or HEK 293 cell line
• Selected mAb producing hybridoma cell line
+ A102: Cloning of Antibody Expression Construct
Cloning of Antibody Expression Construct
Price: $1,200 per sample
Turnaround Time: 10 to 15 (standard pUC-CMV based vectors, time for customer designated vector cloning varies)
Service Description: Cloning of the monoclonal LC gene and HC gene into pUC-CMV based expression vectors or client designated vectors
Deliverables: Two plasmid DNA (20 ug) pUC-CMV based antibody expression constructs, one expressing LC and the other expressing HC
Suitable Sample Type: Product from A101
+ A103: Confirmation of Monoclonal Antibody Sequence
Confirmation of the Amino Acid Sequences of mAbs Using LC-MS/MS
Price: $1,000 per sample
Turnaround Time: 10 to 15 business days
Service Description:
• Amino acid sequences of mAbs are confirmed using liquid chromatography with tandem mass spectrometry
• Workflow: Sample preparation, enzymatic digestion, LC-MS/MS analysis, data analysis, and results report
Deliverables:
• Detailed project report with confirmed amino acid sequence
• List of identified host proteins, if any
• List of identified host proteins with matched peptides sequences
• Purity
Suitable Sample Type: Purified antibody (> 20 µg/sample; purity > 90%)
submission Guidelines
