Creative Biolabs offers the unparalleled mass sequencing service to analyze the T cell receptors (TCR) repertoires. Scientists of Creative Biolabs can provide unbiased TCR repertoire profiling on our Magic™ platform, with minimal loss of quantitative information originally contained in a lymphocyte sample. We are proud to introduce this cutting-edge technology to facilitate the process of immunological research of our global clients.
The Background of Next-generation Sequencing (NGS) for Deep TCR Profiling
The TCR repertoire is a mirror of the human immune system reflecting processes caused by infections, autoimmunity, aging, and diseases. The adaptive immune system drives the immune response via specific hypervariable TCR molecules. This repertoire is generated by a complex series of genetic events. The variable region of each TCR chain consists of three complementary determining regions (CDR) and four frame regions (FR). CDR3 is generated by recombination of variable (V), diversity (D), and joining (J) gene segments in the beta chain, and V and J region gene segments in the alpha chain. High convergence of TCR beta CDR3 amino acid sequences, along with the expanding knowledge on their specificities, suggests the potential for diagnostics of multiple infections and pathological states via deep individual TCR profiling. These perspectives demand the development of powerful approaches for deep unbiased TCR profiling. NGS is a powerful tool for deep TCR profiling. So far, questions abound regarding the methodological approaches for sample preparation and correct data interpretation. Accumulated errors from PCR and sequencing process along with library preparation bottlenecks have resulted in information loss, biased quantification, and generation of huge artificial TCR diversity. Altogether, these technical challenges lead to the loss of the original TCR repertoire of an analyzed T cell sample, generation of false TCR diversity, and inability to interpret sequence information in a quantitative way.
Fig.1 Frequencies of V-J pairing calculated from the subset of clonotypes with an unambiguous TCR beta V gene segment assignment (Freeman et al. 2009).
Unbiased TCR Repertoire Profiling in Creative Biolabs
To solve these problems, Creative Biolabs has developed a powerful approach for unbiased TCR repertoire profiling based on our Magic™ platform. Our scientists have optimized a cDNA-based protocol that allows unbiased pre-sequencing ampliﬁcation of alpha-TCR and beta-TCR gene libraries in the human and murine. The protocol uses speciﬁc primers for cDNA synthesis and introduces sample barcode at the ﬁrst stage of library preparation. This approach allows efﬁcient and unbiased ampliﬁcation of millions of the TCR molecules. In addition, we have developed advanced algorithms for efficient and valid correction of errors with minimal loss of quantitative information. Advanced correction allows the removal of the majority of artificial TCR diversity and rescues most of the sequencing information.
Applications of Our Unbiased TCR Repertoire Profiling Service
With years of research and development experience in the field of immunology, scientists of Creative Biolabs have performed over hundreds of TCR repertoire sequencing projects and accumulated extensive experiences. We can provide unbiased TCR repertoire profiling, allowing the removal of the majority of artificial TCR diversity and rescuing most of the sequencing information. We are pleased to offer the best service with the most accurate results to our global customers.
Please contact us for more information and a detailed quote.