title: Pairing of T-cell receptor chains via emulsion PCR.
abstract: Our ability to analyze adaptive immunity and engineer its activity has long been constrained by our limited ability to identify native pairs of heavy-light antibody chains and alpha-beta T-cell receptor (TCR) chains--both of which comprise coupled "halves of a key", collectively capable of recognizing specific antigens. Here, we report a cell-based emulsion RT-PCR approach that allows the selective fusion of the native pairs of amplified TCR alpha and beta chain genes for complex samples. A new type of PCR suppression technique was developed that makes it possible to amplify the fused library with minimal noise for subsequent analysis by high-throughput paired-end Illumina sequencing. With this technique, single analysis of a complex blood sample allows identification of multiple native TCR chain pairs. This approach may be extended to identify native antibody chain pairs and, more generally, pairs of mRNA molecules that are coexpressed in the same living cells.
authors.affiliation: Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Moscow, Russia.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Turchaninova
authors.forename: Olga V
authors.initials: OV
authors.lastname: Britanova
authors.forename: Dmitriy A
authors.initials: DA
authors.lastname: Bolotin
authors.forename: Mikhail
authors.initials: M
authors.lastname: Shugay
authors.forename: Ekaterina V
authors.initials: EV
authors.lastname: Putintseva
authors.forename: Dmitriy B
authors.initials: DB
authors.lastname: Staroverov
authors.forename: George
authors.initials: G
authors.lastname: Sharonov
authors.forename: Dmitriy
authors.initials: D
authors.lastname: Shcherbo
authors.forename: Ivan V
authors.initials: IV
authors.lastname: Zvyagin
authors.forename: Ilgar Z
authors.initials: IZ
authors.lastname: Mamedov
authors.forename: Carsten
authors.initials: C
authors.lastname: Linnemann
authors.forename: Ton N
authors.initials: TN
authors.lastname: Schumacher
authors.forename: Dmitriy M
authors.initials: DM
authors.lastname: Chudakov
mesh_terms: D001483:Base Sequence; D005784:Gene Amplification; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000032:analysis / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000032:analysis / Q000276:immunology*; D007963:Leukocytes, Mononuclear; D008297:Male; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000276:immunology*; D017422:Sequence Analysis, DNA
chemical_list: D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D016693:Receptors, Antigen, T-Cell, alpha-beta
keywords: Antibodies; Drug design/discovery; Molecular biology; NGS; TCR