![]() VH and VK gene libraries were obtained through subsequent PCR amplification using primers with universal sequences. Double-stranded cDNA was synthesized using primers containing V/J gene–specific regions and universal sequence parts for in vitro display. To minimize this alternation, we devised a new method for harvesting immunoglobulin genes and tested its performance in rabbit variable heavy chain (VH) and variable kappa light chain (VK) genes. However, during this process, the B cell receptor repertoire is distorted due to interactions between multiple target genes and primers. To construct in vitro display antibody libraries from immune repertoires, singleplex or multiplex PCR amplification were employed using primers targeting multiple immunoglobulin genes. Next-generation sequencing analysis confirmed that universal PCR libraries had more diverse VH and VK clonotypes, and a less biased clonal distribution, than conventional singleplex or multiplex gene-specific PCR libraries.ĪB - Immune hosts are valuable sources for antibody discovery. N2 - Immune hosts are valuable sources for antibody discovery. T1 - Amplification of a minimally biased antibody repertoire for in vitro display using a universal primer-based amplification method Next-generation sequencing analysis confirmed that universal PCR libraries had more diverse VH and VK clonotypes, and a less biased clonal distribution, than conventional singleplex or multiplex gene-specific PCR libraries.", Next-generation sequencing analysis confirmed that universal PCR libraries had more diverse V H and V K clonotypes, and a less biased clonal distribution, than conventional singleplex or multiplex gene-specific PCR libraries.Ībstract = "Immune hosts are valuable sources for antibody discovery. V H and V K gene libraries were obtained through subsequent PCR amplification using primers with universal sequences. To minimize this alternation, we devised a new method for harvesting immunoglobulin genes and tested its performance in rabbit variable heavy chain (V H) and variable kappa light chain (V K) genes. Immune hosts are valuable sources for antibody discovery. ![]()
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