The removal of these artifacts will provide an accurate assessment of antibody repertoires and benefit related studies, especially mAb discovery and antibody-guided vaccine design.Īntibodies (Abs), also known as immunoglobulins (Igs), are the most important component of humoral immunity. Tested by ultra-deep Rep-seq data, DUMPArts removed inter-sample chimeras, which cause artifactual shared clones and constitute approximately 15% of reads in the library, as well as intra-sample chimeras with erroneous SHMs and constituting approximately 20% of the reads, and corrected base errors and amplification biases by consensus building. Here, a novel approach named DUMPArts, which improves the accuracy of antibody repertoires by labeling each sample with dual barcodes and each molecule with dual unique molecular identifiers (UMIs) via minimal PCR amplification to remove artifacts, is developed. However, polymerase chain reaction (PCR) amplification introduces extensive artifacts including chimeras and nucleotide errors, leading to false discovery of antibodies and incorrect assessment of somatic hypermutations (SHMs) which subsequently mislead downstream investigations. 8Key Laboratory of Mental Health of the Ministry of Education, Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Southern Medical University, Guangzhou, ChinaĪntibody repertoire sequencing (Rep-seq) has been widely used to reveal repertoire dynamics and to interrogate antibodies of interest at single nucleotide-level resolution.7Division of Nephrology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.6Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.5School of Computer Science and Technology, Beijing Institute of Technology, Beijing, China.4Department of Bioinformatics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.3State Key Laboratory of Organ Failure Research, National Clinical Research Center for Kidney Disease, Division of Nephrology, Nanfang Hospital, Southern Medical University, Guangzhou, China.2Guangdong-Hong Kong Joint Laboratory on Immunological and Genetic Kidney Diseases, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.1Center for Precision Medicine, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China.Qilong Wang 1,2†‡ Huikun Zeng 1,2†‡ Yan Zhu 1,2†‡ Minhui Wang 3†‡ Yanfang Zhang 3,4† Xiujia Yang 3,4† Haipei Tang 1,2† Hongliang Li 5 Yuan Chen 1,2† Cuiyu Ma 4† Chunhong Lan 1,2,3,4† Bin Liu 5 Wei Yang 6*† Xueqing Yu 2,7* Zhenhai Zhang 1,2,3,4,8*†
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