Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes

被引:11
作者
Devulapally, Praneeth Reddy [1 ]
Buerger, Joerg [2 ,3 ]
Mielke, Thorsten [2 ]
Konthur, Zoltan [4 ]
Lehrach, Hans [5 ,6 ]
Yaspo, Marie-Laure [1 ,5 ]
Gloekler, Joern [7 ]
Warnatz, Hans-Joerg [1 ]
机构
[1] Max Planck Inst Mol Genet, Otto Warburg Lab Gene Regulat & Syst Biol Canc, Berlin, Germany
[2] Max Planck Inst Mol Genet, Microscopy & Cryoelectron Microscopy Serv Grp, Berlin, Germany
[3] Charite, Inst Med Phys & Biophys, Berlin, Germany
[4] Max Planck Inst Colloids & Interfaces, Dept Biomol Syst, Potsdam, Germany
[5] Alacris Theranost GmbH, Berlin, Germany
[6] Dahlem Ctr Genome Res & Med Syst Biol, Berlin, Germany
[7] Tech Univ Appl Sci Wildau, Inst Appl Biosci, Dept Mol Biotechnol & Funct Genom, Wildau, Brandenburg, Germany
来源
GENOME MEDICINE | 2018年 / 10卷
关键词
MEMORY B-CELLS; IMMUNOLOGICAL REPERTOIRE; MONOCLONAL-ANTIBODIES; IMMUNOGLOBULIN HEAVY; GENE REPERTOIRE; EMULSION PCR; PLASMA-CELLS; EXPRESSION; IDENTIFICATION; TRANSLATION;
D O I
10.1186/s13073-018-0542-5
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Existing methods for paired antibody heavy- and light-chain repertoire sequencing rely on specialized equipment and are limited by their commercial availability and high costs. Here, we report a novel simple and cost-effective emulsion-based single-cell paired antibody repertoire sequencing method that employs only basic laboratory equipment. We performed a proof-of-concept using mixed mouse hybridoma cells and we also showed that our method can be used for discovery of novel antigen-specific monoclonal antibodies by sequencing human CD19(+) B cell IgM and IgG repertoires isolated from peripheral whole blood before and seven days after Td (Tetanus toxoid/Diphtheria toxoid) booster immunization. We anticipate broad applicability of our method for providing insights into adaptive immune responses associated with various diseases, vaccinations, and cancer immunotherapies.
引用
收藏
页数:12
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