14 Finally, filtering and error rate assessment should be performed with extreme caution

because the rare receptor sequences that are presented at very low levels in an individual might be mistaken for error-containing sequences and ignored. Construction of synthetic antibody libraries is important for therapeutic antibody development. Recent studies have presented novel methods for library design combining NGS. These libraries are generated by introducing diversity in the variable region of the antibody43,44 and high throughput sequencing was used to characterize the coverage and diversity of VH and Vκ sequences. High throughput sequencing analysis can also be used for the production of mono-clonal antibodies. Massive production of antigen-specific antibodies is essential for both research and clinical aspects, mainly for diagnostic APO866 cost and therapeutic treatments (cancer, autoimmune diseases etc.). Reddy et al.

used massively parallel sequencing technology Selleck Veliparib for antibody isolation to overcome the extremely time-consuming step of screening for recombinant antibodies that was used previously.45 Recombinant genes are synthesized from paired VH and VL segments, based on the understanding that VH and VL have relatively similar expression frequencies and originate from the same B cell, and therefore constitute the complete antibody dimer. Large-scale sequencing of rearranged immune receptor genes can also be of use in the detection and tracking of clonally expanded B-cell and T-cell populations in different physiological and pathological conditions. Lymphocyte malignancies usually originate Orotic acid from a single dominant immunoglobulin or TCR. Therefore, obtaining information about the relative abundance of these receptors using high throughput sequencing methods might be key for better understanding their nature. Large-scale sequencing of the immune receptors repertoire offers distinct and highly detailed molecular characterization that

may reform our perception of the immune system while supporting diagnosis, prognosis and monitoring of disease. Ever since its introduction as a well-established method only a few years ago, NGS has emerged as a major player in molecular biology, genomics, systems biology and other fields.46 Next-generation sequencing promises to make a similar impact in immunity, and presents, for the first time, an opportunity for a comprehensive view of the T-cell and B-cell repertoires. As much as this technology presents an opportunity, it brings with it major challenges in data storage and data analysis. We need to consider human ability to store these data, to view these data and to produce meaning from the data. The community’s interest in sequencing and its applications promises some of the solutions as already available.