RT Journal Article
SR Electronic
T1 Antibody Upstream Sequence Diversity and Its Biological Implications Revealed by Repertoire Sequencing
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.09.02.280396
DO 10.1101/2020.09.02.280396
A1 Yan Zhu
A1 Xiujia Yang
A1 Jiaqi Wu
A1 Haipei Tang
A1 Qilong Wang
A1 Junjie Guan
A1 Wenxi Xie
A1 Sen Chen
A1 Yuan Chen
A1 Minhui Wang
A1 Chunhong Lan
A1 Lai Wei
A1 Caijun Sun
A1 Zhenhai Zhang
YR 2020
UL http://biorxiv.org/content/early/2020/09/03/2020.09.02.280396.abstract
AB The sequence upstream of antibody variable region (Antibody Upstream Sequence, or AUS) consists of 5’ untranslated region (5’ UTR) and two leader regions, L-PART1 and L-PART2. The sequence variations in AUS affect the efficiency of PCR amplification, mRNA translation, and subsequent PCR-based antibody quantification as well as antibody engineering. Despite their importance, the diversity of AUSs has long been neglected. Utilizing the rapid amplification of cDNA ends (5’RACE) and high-throughput antibody repertoire sequencing (Rep-Seq) technique, we acquired full-length AUSs for human, rhesus macaque (RM), cynomolgus macaque (CM), mouse, and rat. We designed a bioinformatics pipeline and discovered 2,957 unique AUSs, corresponding to 2,786 and 1,159 unique sequences for 5’ UTR and leader, respectively. Comparing with the leader records in the international ImMunoGeneTics (IMGT), while 529 were identical, 313 were with single nucleotide polymorphisms (SNPs), 280 were totally new, and 37 updated the incomplete records. The diversity of AUSs’ impact on related antibody biology was also probed. Taken together, our findings would facilitate Rep-Seq primer design for capturing antibodies comprehensively and efficiently as well as provide a valuable resource for antibody engineering and the studies of antibody at the molecular level.Competing Interest StatementThe authors have declared no competing interest.