Compared to hybridoma technology that retains the natural paring of the heavy and light chains, display technologies allow random pairing of the heavy and light chains, which generates additional antibody diversity. Following the invention of phage display technology, various other display technologies, such as yeast display 13 and ribosome display 14, emerged, which further enhanced rapid discovery of antibodies. Phage display is now one of the most widely used technologies for antibody discovery and engineering 12. It was also the first approved human mAb and is currently the best-selling antibody drug on the market 11. In 2002, adalimumab became the first phage display-derived antibody approved by FDA 10. The invention of polymerase chain reaction (PCR) in 1983 and phage display technology in 1985 further enabled batch amplification of antibody genes and isolation of target-specific binding antibodies from combinatorial antibody libraries in a high-throughput manner, which created a paradigm shift in antibody drug discovery 8, 9. The invention of recombinant DNA technology in the early 1970’s allowed gene cloning and subsequently protein engineering for desired properties 3, 4, 5, 6, 7. The first monoclonal antibody (mAb), muromonab-CD3 (OKT3), approved by FDA in 1986 was derived from hybridoma screening 2. The invention of hybridoma technology in 1975 led to the first paradigm in antibody drug discovery 1. The history of antibody drug discovery reflects the advancement of antibody screening technology. This new method may create a new paradigm in antibody drug discovery. In contrast, we obtained only neutral binders and antibody antagonists from the same repertoire by phage display, suggesting that the new approach described here is more efficient than traditional methods in isolating functional antibodies. We validated this method by identifying a panel of antibody antagonists and an antibody agonist to the human apelin receptor from an immune antibody repertoire. This method links antibody genotype with phenotype and is applicable to all GPCR targets. ![]() Here we describe a function-based high-throughput screening method for quickly identifying antibody antagonists and agonists against GPCRs by combining glycosylphosphatidylinositol-anchored antibody cell display with β-arrestin recruitment-based cell sorting and screening. However, for complex membrane proteins, such as G protein-coupled receptors (GPCRs), binding-based screening rarely results in functional antibodies. Hybridoma and phage display are two powerful technologies for isolating target-specific monoclonal antibodies based on the binding.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |