Immune hosts are valuable sources for antibody discovery. To construct in vitro display antibody libraries from immune repertoires, singleplex or multiplex PCR amplification were employed using primers targeting multiple immunoglobulin genes. However, during this process, the B cell receptor repertoire is distorted due to interactions between multiple target genes and primers. 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. Double-stranded cDNA was synthesized using primers containing V/J gene–specific regions and universal sequence parts for in vitro display. V_H and V_K gene libraries were obtained through subsequent PCR amplification using primers with universal sequences. 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.

免疫宿主是抗体发现的宝贵来源。为了从免疫组库构建体外展示抗体文库，使用针对多个免疫球蛋白基因的引物采用单重或多重 PCR 扩增。然而，在此过程中，由于多个靶基因和引物之间的相互作用，B 细胞受体库会发生扭曲。为了最大限度地减少这种变化，我们设计了一种新的免疫球蛋白基因采集方法，并测试了其在兔可变重链 (V _H ) 和可变κ轻链 (V _K ) 基因中的性能。使用含有 V/J 基因特异性区域和通用序列部分的引物合成双链 cDNA，用于体外展示。V _{^ h}和V _K基因文库通过随后使用具有通用序列的引物进行PCR扩增获得。新一代测序分析证实，与传统的单重或多重基因特异性 PCR 文库相比，通用 PCR 文库具有更多样化的 V _H和 V _K克隆型，且克隆分布的偏向性更小。