Antibody discovery technology: innovation and outlook from classic to leading edge

Antibodies, as core tools in life science research and clinical therapeutics, have witnessed revolutionary advancements in discovery technologies—evolving from the traditional hybridoma technique to modern high-throughput and intelligent platforms. This review systematically summarizes major progress in antibody discovery technologies. It begins with an elaboration of the foundational hybridoma technology, including its principles, advantages, and limitations, along with improvement strategies such as humanization. The article then shifts to various in vitro display technologies—such as phage, yeast, and ribosome display—with a focus on their core mechanisms, library construction, and screening strategies. The review further elaborates on the advantages and implementation approaches of single B-cell antibody technology, emphasizing its critical role in preserving natural heavy-light chain pairing information. Also explored are applications of next-generation sequencing in antibody repertoire analysis and dynamic tracking, as well as the cutting-edge role of computational biology and artificial intelligence in antibody design, optimization, and developability prediction. Additional topics include strategies for challenging targets, transgenic animal platforms, engineering of bispecific and multispecific antibodies, and the potential of synthetic biology in novel antibody development. Finally, by comparing key metrics across different technological platforms, the review outlines current challenges and anticipates future directions, including the integration of multiple technologies, AI-enabled development, and personalized antibody therapeutics. This comprehensive review provides technical insights and strategic guidance for antibody drug R&D, offering valuable reference for promoting innovation and clinical application of therapeutic antibodies.