Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing

AbstractAntibodies are immune system proteins that recognize noxious molecules for elimination. Their sequence diversity and binding versatility have made antibodies the primary class of biopharmaceuticals. Recently it has become possible to query their immense natural diversity using next-generation sequencing of immunoglobulin gene repertoires (Ig-seq). However, Ig-seq outputs are currently fragmented across repositories and tend to be presented as raw nucleotide reads, which means nontrivial effort is required to reuse the data for analysis. To address this issue, we have collected Ig-seq outputs from 53 studies, covering more than half a billion antibody sequences across diverse immune states, organisms and individuals. We have sorted, cleaned, annotated, translated and numbered these sequences and make the data available via our Observed Antibody Space (OAS) resource at antibodymap.org. The data within OAS will be regularly updated with newly released Ig-seq datasets. We believe OAS will facilitate data mining of immune repertoires for improved understanding of the immune system and development of better biotherapeutics.

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The Application of Next Generation Sequencing to the Understanding of Antibody Repertoires

Frontiers in Immunology ◽

10.3389/fimmu.2013.00265 ◽

2013 ◽

Vol 4 ◽

Cited By ~ 23

Author(s):

Pascale Mathonet ◽

Christopher G. Ullman

Keyword(s):

Next Generation Sequencing ◽

Next Generation ◽

Antibody Repertoires ◽

Generation Sequencing

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Deciphering antigen-responding antibody repertoires by using next-generation sequencing and confirming them through antibody-gene synthesis

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2017.04.054 ◽

2017 ◽

Vol 487 (2) ◽

pp. 300-306 ◽

Cited By ~ 8

Author(s):

Naoko Kono ◽

Lin Sun ◽

Hiroyuki Toh ◽

Takeyuki Shimizu ◽

Hanbing Xue ◽

...

Keyword(s):

Next Generation Sequencing ◽

Gene Synthesis ◽

Next Generation ◽

Antibody Repertoires ◽

Generation Sequencing ◽

Antibody Gene

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Development of a potent and protective germline-like antibody lineage against Zika virus in a convalescent human

10.1101/661918 ◽

2019 ◽

Author(s):

Fei Gao ◽

Xiaohe Lin ◽

Linling He ◽

Ruoke Wang ◽

Han Wang ◽

...

Keyword(s):

Next Generation Sequencing ◽

Zika Virus ◽

Neutralizing Antibodies ◽

Great Promise ◽

Human Antibody ◽

Next Generation ◽

Zikv Infection ◽

Variable Regions ◽

Next Generation Sequencing Ngs ◽

Generation Sequencing

AbstractZika virus (ZIKV) specific neutralizing antibodies hold a great promise for antibody-based interventions and vaccine design against ZIKV infection. However, their development in infected patients remain unknown. Here, we report on the dynamic development of a potent and protective ZIKV-specific human antibody ZK2B10 initially isolated from a ZIKV convalescent individual using next-generation sequencing (NGS). The unbiased repertoire analysis showed dramatic changes in many families of heavy and light chain variable regions. However, lineage tracing of ZK2B10 revealed limited somatic hypermutation throughout the 12 months since the onset of symptom. In particular, NGS-derived germline-like somatic variants neutralized and protected mice from lethal challenge of ZIKV without detectable cross-reactivity with Dengue virus (DENV). Site-directed mutagenesis identified two residues within λ chain, N31 and S91 that are essential to the functional maturation. The dynamic features unveiled here will assist us to better understand the pathogenesis of ZIKV infection and inform rational design of vaccines.Author summaryRecently emerged ZIKV is associated with severe neurological complications such as Guillain–Barré syndrome in adults and congenital microcephaly in newborns. No ZIKV-specific therapeutics or vaccines are currently available. We and others have identified a number of neutralizing antibodies capable of protecting experimental animals from ZIKV infection. However, the development of these potent antibodies during ZIKV natural infection remains unknown. Here, we report on the longitudinal analysis of one such antibody ZK2B10 using next-generation sequencing (NGS), bioinformatics and functional analysis. We found that the ZK2B10 germline-like antibodies possess strong neutralizing activity in vitro and impressive protectivity against lethal ZIKV infection in vivo. These findings suggest that the potent and protective antibody response against ZIKV can be generated within relative short term with high germline identity which provide great hope and promise for successful vaccine development against ZIKV.

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