With the efforts of its scientists and years of experience in the field of antibody engineering, Creative Biolabs has conquered the current shortcomings of sequencing based on traditional methods and self-developed “Database Assisted Shotgun Sequencing” (DASS) technology, which provides de novo antibody sequencing service for research, diagnostic and therapeutic industries.
This novel service consists of variable region, variable plus leader region, and full length heavy and light chain antibody sequencing for all species. Purified monoclonal antibodies in multivalent forms can be sequenced with 100% coverage of the desired regions as well as excellent accuracy. “Apart from that, all species, isotypes and allotypes of antibodies can also be sequenced with 100% coverage.” said Monica Müller, scientific officer of Creative Biolabs.
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 introduces hypermutations into the sequence so as to optimize the affinity. Creative Biolabs’ mapping algorithm is error tolerant and can match the “mutated” peptides to the corresponding germline.
Sequencing of isobaric amino acids
Compared with other MS based methods, Creative Biolabs can discriminate most isobaric amino acid combinations. W can be distinguished from GE, AD and SV; R can be distinguished from GV; Q can be distinguished from GA and K; Leucine and isoleucine cannot be distinguished. However, most of these positions can be determined using the corresponding germline sequence.
De Novo Sequence of the CDR3 region
While the CDR3 of the light chain is mostly encoded by the germline sequences, the CDR3 of the heavy chain is usually not available in databases. It is encoded by the so called D-segments, but these are modified by nucleases and terminal transferases. Basicly, only 1-4 AA of a D-segment remain in the matured antibody. The rest of the D-segment is “artificial” and has to be sequenced de Novo. Creative Biolabs’ method generates many overlapping peptides during the fragmentation process, enabling it to sequence very long stretches of unknown amino acids. “The technique is so powerful that we are able to sequence a 20 kDa protein, which had no homologue in the database.”
About Creative Biolabs
Creative Biolabs is specialized in providing custom biotechnology and pharmaceutical services that cover the full scope of biotechnology needs of early antibody development and drug discovery. As a trusted provider of the most cost-effective outsourcing solutions, Creative Biolabs has been working for a large number of clients from biotechnology and pharmaceutical companies as well as government and academic research laboratories all over the world.
Company Name: Creative Biolabs
Contact Person: Bella Smith
Email: Send Email
Address:45-1 Ramsey Road
State: New York
Country: United States