Accurately acquiring antibody sequences is one of the most important prerequisites to achieving antibody functional optimization and expression of recombinant engineering antibodies. Antibody sequencing and analysis also plays a key role for researchers publishing articles and apply for patents. The data is even more critical for pharmaceutical enterprises to further their research and develop of novel drugs. Synbio Technologies can quickly and accurately sequence and analyze antibody sequences with our advanced GenoTM Ab Platform. Using this platform, we can perform high quality de novo sequencing of multi-species’ (including mice, rats, Armenian hamsters, camels, rabbits, equines, bovines, canines, birds, et al.) monoclonal antibodies. The Syno® Ab Platform also has the capabilities to analyze data that may come from various antibody libraries and hybridoma technologies, making this platform very applicable. Our experienced bioinformatics analysis team can also provide professional data mining and annotation.
Synbio Technologies can utilize immune repertoire sequencing technology to selectively amplify the target region of T/B lymphocytes. This approach combined with high-throughput sequencing technology and powerful antibody big data platform. In doing so, we can comprehensively evaluate the diversity of the immune system. The combination of these technologies has been widely used in the rapid antibody preparation, disease and immune system surveillance, as well as vaccine optimization. In addition to providing the genetic sequences of the monoclonal antibody, we can also express and purify antibodies by utilizing our NGTM Codon Optimization Technology and Syno® DNA Synthesis Platform.
- We use custom primer sets and special reagents developed in-house to ensure high success rates.
- Our unique internal data mining tool can remove redundancies and extract information of target antibody sequence.
- Diversified NGS analysis platform has the powerful ability for antibody gene analysis, resulting in a high quality output.
- Complete bioinformatics analysis removes redundant sequence data, and deeply mining the high-value information.
Standard Antibody Sequencing Service Process
Synbio Technologies’ Immune repertoire sequencing and bioinformatics analysis:
- Library Construction
Synbio Technologies performed 5’ RACE reverse transcription of the extracted RNA, amplified and constructed the library.
- Library Quality Control
The Qubit 2.0 Fluorometer was used to perform preliminary quantification after the library construction. After dilution, insert size detection was performed on the library based on the Agilent 2100 bioanalyzer. Only when meeting expected requirements, the effective concentration of the library (>2nM) was accurately quantified by qPCR to ensure the quality of the library.
- Library Sequencing
Different libraries were pooled and sequenced by Illumina Miseq/HiSeq according to the effective concentration and the required amount of target data. Four fluorescent labeled dNTPs, DNA polymerase and adaptors were added to the flow cell for the amplification. When each sequence cluster extended and generated the complementary chain, the corresponding fluorescent signals were released and captured after each new dNTP was added. The optical signals were then transformed into sequencing peaks by professional software to obtain the sequence information of the fragments.
- Data Analysis
Quality assessment of sequencing data.