Recent years, the biotechnology industry maintains the trend of rapid developing. In a series of biotechnology applications, some basic techniques and devices are used in almost every case, such as liquid pipettng, cell treatment, sample preparation and so on. While speaking of these applications, we can never miss the next generation sequencing technology. Following the appearance and widely use of the new generation sequencing technology, biological drugs like monoclonal antibodies and other biopharmaceutical production are gradually occupying the significant position of the pharmaceutical industry.
Based on traditional methods and developed “Database Assisted Shotgun Sequencing” (DASS) technology, Creative Biolabs has overcome the current drawback of sequencing so as to provide better de novo antibody sequencing service for researchers, diagnostic and therapeutic industries.
Its services consist of variable region, variable plus leader region, and full length heavy and light chain antibody sequencing for all species. All species, including isotypes and allotypes of antibodies, can be sequenced with 100% coverage. Numerous successful cases from Creative Biolabs have proved that it can meet customers’ needs by offering antibody sequencing with 100% accuracy.
Service Specifications—Sequencing of the V and J and C segments by DASS
The V and J and C gene segments of antibodies are available in public databases. However, during the maturation of an antibody, the B-cell will introduce hypermutations into the sequence to optimize the affinity. Our mapping algorithm is error tolerant and can match the “mutated” peptides to the corresponding germline accurately.
Because of the large number of peptides, Creative Biolabs usually gets sequence information for every peptide bond in the antibody. Typically, 20-70 different MS/MS spectra are generated for each amino acid (AA) position. Hence, even the hardest sequences of peptides with rich proline and arginine can be resolved. As the order of all amino acids is clear, there is no need of time consuming techniques like edman sequencing.
Creative Biolabs also identifies the closest allele in the IMGT database for a light chain.
Before starting the sequencing, the masses of the reduced light and heavy chain will be determined. At this step scientists from Creative Biolabs see if your mAB contains a nonsense light chain, as the corresponding masses are known. The masses of the intact chains serve to validate the determined sequences. The light chain sequence must match within a +/- 1.2 Da range. For the heavy chain +/- 1.7 Da are allowed. If the determined sequences do not match with the protein mass, you will not be charged.
In addition, the DASS system above provides reliable supporting proof for confirmation and validation of produced mAb, QC analysis and IND/NDA application.