Yeast Display Library Construction Introduction

In recent years, yeast surface display has proved to be a versatile technology for use in the biomedical field, and it is important in areas such as antibody library screening, and antibody and protein engineering. However, the construction of yeast display libraries is a tedious and time-consuming process. It involves the generation of heavy and light chain diversity in haploid yeast cells that need to be bound by yeast mating. Therefore, the proper construction of yeast display libraries is the basis for subsequent applications. Based on our rich field experience and yeast display platform, Creative Biolabs provides comprehensive services to support the development and application of yeast display library construction.

Yeast Display Library Construction

Yeast display technology plays an important role in areas such as antibody and protein engineering. For example, yeast display-based T cell receptor engineering has been used for the development of TCR antibody drugs and cell therapies. In antibody engineering, yeast display technology has been widely used for human monoclonal antibody identification, antibody screening and optimization. However, yeast display library construction and yeast display library screening are important steps in yeast display technology, which are crucial to ensure the success of subsequent studies.

There are two common methods of library construction. One is SMART amplification technology and another is Gateway recombination technology. In actual research, the specific choice of which method needs to be determined based on the amount of starting RNA. SMART technology, i.e., RNA template 5' end conversion mechanism. It can continuously synthesize sequences in the 5' untranslated region, which contains a larger proportion of full-length clones. Specifically, it refers to the co-transformation of yeast by a synthesized cDNA and a linearized vector, where the cDNA and vector are ligated to form a complete plasmid in vivo by the action of recombinase. The cDNA library constructed by SMART technology is representative of the abundance of mRNA in the original sample, preserving the integrity of the biogenetic information. Gateway technology, which utilizes proprietary recombinant sequences, allows for more efficient transfer of DNA fragments into the plasmid. This method can be applied to cloning large fragments of genes and allows for the transfer of DNA between different expression vectors while maintaining the correct reading frame. In the process of library construction using Gateway technology, no PCR amplification is required, so a high amount of starting RNA is required, which is more than 300 μg. Under the condition of sufficient sample volume, yeast libraries constructed by the Gateway method avoided the effects of PCR amplification on the libraries, such as the loss of low-abundance genes and the possible mutation of individual bases during the amplification process.

Creative Biolabs has a wealth of knowledge and experience in yeast display. We would be happy to share with you our knowledge and experience on yeast display library construction.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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