Antibody upstream sequence diversity and its biological implications revealed by repertoire sequenci

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The sequence upstream of the antibody variable region (antibody upstream sequence[AUS]) consists of a 5\'untranslated region (5\'UTR) and a preceding leader region.The sequence variations in AUS affect antibody engineering end PCR based antibody quantification and may also be implicated in mRNA tran-scription and translation.However,the diversity of AUSs remains elusive.Using 5\'rapid amplification of cDNA ends and high-throughput antibody repertoire sequencing technique,we acquired full-length AUSs for human,rhesus macaque,cynomolgus macaque,mouse,and rat.We designed a bioinformatics pipeline and identified 3307 unique AUSs,corresponding to 3026 and 1457 unique sequences for 5\'UTR and leader region,respectively.Comparative analysis indicated that 928 (63.69%) leader sequences are novel relative to those recorded in the international ImMunoGeneTics information system.Evolutionarily,leader se-quences are more conserved than 5\'UTR and seem to coevolve with their downstream V genes.Besides,single-nucleotide polymorphisms are position dependent for leader regions and may contribute to the functional reversal of the downstream V genes.Finally,the AUGs in AUSs were found to have little impact on gene expression.Taken together,our findings can facilitate primer design for capturing antibodies efficiently and provide a valuable resource for antibody engineering and molecule-level antibody studies.
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