The accurate measurement of the immune repertoire is important for understanding the repertoire response in infections, vaccinations and cancer immunology. Creative Biolabs has established a high-throughput Magic™ sequencing platform for large-scale sequencing services. Our scientists developed an advanced approach to defining the immunoglobulin repertoire of feline by analyzing complementarity determining region 3 (CDR3) of the heavy chain. The data will provide signiﬁcant insight into understanding the feline immune system function.
The development of next-generation sequencing (NGS) has made antibody repertoire sequencing feasible and affordable. The proﬁling of entire antibody repertoires offers new possibilities for studying whole antibody repertoires of individuals. These sequencing approaches, combined with bioinformatics, allow us to study and compare individuals. It also allows the precise determination of entire antibody lineages. In the last decades, increasing interest has been aroused in the cat as a model for speciﬁc diseases. The feline immune system, similar to canine or human immune system, generates antibody diversity by V (D) J recombination. In the recombination process, a diverse assortment of variable genes (V), diversity (D) genes and joining (J) genes are recombined to create an antibody variable domain. Further post-recombination processes like insertion and deletion of nucleotides and somatic hypermutation can considerably increase the diversity of the variable region. Felines show the mammalian organization of the IGH genome and the IG locus has been described previously. Domestic cats (Felis catus) have been studied in detail immunologically; however, the actual, expressed feline antibody repertoire has not been characterized. Although Felis catus has the same IGHV gene structure as Canidae family, the feline IGHV locus is largely devoid of orthologs. Simultaneously, they are an unrivaled platform for engineering highly effective diagnostic and therapeutic reagents for companion animals.
Fig.1 CDR-H3 Diversity IgG (Steiniger et al. 2017).
Creative Biolabs has developed an effective strategy for the comprehensive characterization of the feline immunoglobulin repertoire. The procedure mainly consists of (1) isolation and quantitation of total RNA, (2) cDNA synthesis and generation of feline IGH and IGL amplicons, and (3) deep sequencing of feline VH and VL gene repertoires on our high-throughput Magic™ BCR repertoire analysis platform. The bioinformatics analysis includes gene usage, length distribution of VH and CDR amino acid distribution and variability by position. With this stringency, poorer quality sequences will not merge and all non-merging paired reads will be discarded. This strategy is an important channel for the identification of high-afﬁnity and feline antibodies.
Key advantages including but not limited to:
With years of research and development experience in the field of immunology, Creative Biolabs has owned a group of seasoned scientists. We can provide the high-quality service for the comprehensive characterization of feline immunoglobulin repertoire. We are pleased to share our extensive experiences with our global clients and facilitate their meaningful immune repertoire research.
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