Role of messenger RNA–ribosome complex in complementary DNA display

A High Performance Platform Based on cDNA Display for Efficient Synthesis of Protein Fusions and Accelerated Directed Evolution

“…In the recent past, there have been notable developments in cDNA display technology in terms of time required for the execution of each round of cDNA display/selection, improvements in ligation reactions, restriction digestion, and increased purified fusions for higher library size. ,− The progress in the faster execution of ligation reactions from 1 h to 10 min to 30 s is highly notable; however, the use of ribonucleases and endonucleases should be avoided to refrain from any sort of contamination that may negatively affect the whole selection process. ,, These methods require 2–3 days/round. In a previous report, the purification steps have been modified to increase the efficiency of the system by reducing the time of processing to 2 days/round .…”

“…Alternatively, custom designed libraries that do not contain stop codons should be used to counter this problem; however, it is noteworthy that custom designed libraries are expensive to prepare. Furthermore, the untranslated mRNA-linker present in the unpurified mixtures (∼70–80%) will participate in the selection process by the formation of aptamers and thus interfere with the outcome of selection. ,, Recently, cDNA display was also improved by the introduction of purification protocols that increased the fusions by approximately 10-fold that can yield libraries by a higher order . However, this method requires an execution time of 2–3 days/round.…”

“…Thus, we expect this cDNA display method to provide higher diversity libraries for searching novel ligands at reduced cost, time, and labor and to combine the intrinsic throughput with high-throughput systems to accelerate the discovery process. The recent developments, including the optimized linkers and methods, ,− and higher-throughput systems , are expected to provide necessary support and significantly impact discovery research.…”

“…11,12,20 Recently, cDNA display was also improved by the introduction of purification protocols that increased the fusions by approximately 10-fold that can yield libraries by a higher order. 37 However, this method requires an execution time of 2−3 days/round.…”