IMGT®,国际ImMunoGeneTics信息系统®(http://www.imgt.org),于1989年在蒙彼利埃创建,法国(CNRS和蒙彼利埃大学)管理抗原受体的巨大和复杂的多样性,是免疫信息学的起源,免疫遗传学和生物信息学之间的一门科学。免疫球蛋白(IG)或抗体和T细胞受体(TR)在IMGT®数据库和工具中的受体水平进行管理和描述。链和域。通过IMGT/V-QUEST(自1997年以来在线,每批50个序列)进行IG和TR可变(V)结构域重排核苷酸序列的分析,对于下一代测序(NGS),IMGT/HighV-QUEST,IMGT/V-QUEST的高通量版本(门户始于2010年,每批500,000个序列)。广泛筛选模拟免疫适应性应答的体内天然多样性的工程化抗体单链片段变量(scFv)的体外组合文库,以发现新的抗原结合特异性。然而,NGS全长scFv(~850bp)的分析代表了一个挑战,因为它们含有通过接头连接的两个V结构域,并且没有用于分析单链中的两个V结构域的工具。
功能“单链片段变量(scFv)分析”已在IMGT/V-QUEST中实现,对于NGS,在IMGT/HighV-QUEST中分析IG和TRscFv的两个V结构域。它分为五个步骤:搜索第一个最接近的V-REGION,第一个V-(D)-J区域的完整表征,然后搜索第二个V区和第二个V-(D)-J区的完整特征,最后是连接体的定界。
对于每个序列或NGS读数,5'V域的位置,scFv中的接头和3'V-结构域以''V-取向的''意义提供。每个V-域都有充分的特征(基因识别,序列描述,连接分析,突变和氨基变化的表征)。功能是通用的,可以分析含有两个V结构域的任何IG或TR单链核苷酸序列,前提是相应的物种IMGT参考目录可用。
在IMGT/V-QUEST中实现的“单链片段变量(scFv)分析”,对于NGS,IMGT/HighV-QUEST中提供了来自组合文库的全长scFv(〜850bp)核苷酸序列的两个V结构域的鉴定和完整表征。还可以对表达的抗原受体IG或TR库的串联配对链进行分析。
IMGT®, the international ImMunoGeneTics information system® ( http://www.imgt.org ), was created in 1989 in Montpellier, France (CNRS and Montpellier University) to manage the huge and complex diversity of the antigen receptors, and is at the origin of immunoinformatics, a science at the interface between immunogenetics and bioinformatics. Immunoglobulins (
IG) or antibodies and T cell receptors (TR) are managed and described in the IMGT® databases and tools at the level of receptor, chain and domain. The analysis of the
IG and TR variable (V) domain rearranged nucleotide sequences is performed by IMGT/V-QUEST (online since 1997, 50 sequences per batch) and, for next generation sequencing (NGS), by IMGT/HighV-QUEST, the high throughput version of IMGT/V-QUEST (portal begun in 2010, 500,000 sequences per batch). In vitro combinatorial libraries of engineered antibody single chain Fragment variable (scFv) which mimic the in vivo natural diversity of the immune adaptive responses are extensively screened for the discovery of novel antigen binding specificities. However the analysis of NGS full length scFv (~850 bp) represents a challenge as they contain two V domains connected by a linker and there is no tool for the analysis of two V domains in a single chain.
The functionality \"Analyis of single chain Fragment variable (scFv)\" has been implemented in IMGT/V-QUEST and, for NGS, in IMGT/HighV-QUEST for the analysis of the two V domains of
IG and TR scFv. It proceeds in five steps: search for a first closest V-REGION, full characterization of the first V-(D)-J-REGION, then search for a second V-REGION and full characterization of the second V-(D)-J-REGION, and finally linker delimitation.
For each sequence or NGS read, positions of the 5\'V-DOMAIN, linker and 3\'V-DOMAIN in the scFv are provided in the \'V-orientated\' sense. Each V-DOMAIN is fully characterized (gene identification, sequence description, junction analysis, characterization of mutations and amino changes). The functionality is generic and can analyse any
IG or TR single chain nucleotide sequence containing two V domains, provided that the corresponding species IMGT reference directory is available.
The \"Analysis of single chain Fragment variable (scFv)\" implemented in IMGT/V-QUEST and, for NGS, in IMGT/HighV-QUEST provides the identification and full characterization of the two V domains of full-length scFv (~850 bp) nucleotide sequences from combinatorial libraries. The analysis can also be performed on concatenated paired chains of expressed antigen receptor
IG or TR repertoires.