The Complete Guide to the Phage Display Technology

1. 2019 The Complete Guide to the Phage Display Technology SCIENCE

2. CONTENTS Part2 Part 1 Part3 Part4 Background Principle Common Methods Profile

3. Part 1 Background of Phage Display Technology

4. Stephen Grant Smith Sir Gregory Paul Winter • George Pearson Smith (born March 10, 1941) is an American biologist and Nobel laureate. He is a Curators' Distinguished Professor Emeritus of Biological Sciences at the University of Missouri in Columbia, Missouri, US.

5. BACKGROUND On October 30, 2018, the Royal Swedish Academy of Sciences announced that half of the 2018 Nobel Prize in Chemistry would be awarded to American scientist G. Smith and British scientist Sir G. Winter in recognition of their contribution to the invention and application of phage display technology.

6. DEVELOPMENT 1.Development of Phage Display Technology Literally speaking, bacteriophages refer to viruses that can "eat" bacteria, i.e. a group of viruses that can infect bacteria, fungi, actinomycetes and spirochetes that are very small in size with morphology varied from species to species. Generally, bacteriophages can be tadpole-shaped, small spherical or slender rod-shaped. Most of the bacteriophages are tailed diohedral structures, while some are not. The capsid proteins regularly arrange to form the head of the icosahedron containing the genetic material - nucleic acid. The filamentous bacteriophage is linear in shape and has no obvious head structure. Its capsid protein particles form a spiral structure, which encapsulates the nucleic acid of the bacteriophage. • Development of Phage Display Technology

7. DEVELOPMENT At present, according to the phage proteomic tree taxonomy, bacteriophages can be classified into 8 categories: smooth bacteriophage, cystic bacteriophage, filamentous bacteriophage, microphage, short tail bacteriophage, long tail bacteriophage, muscle tail bacteriophage and Fuselloviridi. Federick William Twort, a British microbiologist, firstly discovered the phenomenon that bacteriophage can kill bacteria in the culture of Staphylococcus aureus in 1915. As a natural enemy of bacteria, bacteriophages are quickly adopted in the treatment of bacterial infections. In 1919, bacteriophage therapy successfully cured childhood dysentery in France. Later, it was found that bacteriophages can infect many pathogenic bacteria, such as Vibrio cholerae and Bacillus anthracis, and can lyse these host pathogenic bacteria. It has become a specific drug for the treatment of these pathogenic bacteria infection-related diseases and one of the effective methods for anti-infection treatment. With the invention and use of antibiotics, the use of bacteriophages for the treatment of bacterial infections was once at a low ebb. But since the 21st century, due to the bacterial resistance to antibiotics has become more and more serious, bacteriophage therapy once again entered the field of vision, and has achieved preliminary success in the treatment of drug-resistant bacteria and conditional pathogenic bacterial infections. • Development of Phage Display Technology

8. Part 2 Principle of Phage Display Technology

9. Infographic Genes encoding signal peptide of capsid protein and genes encoding capsid protein polypeptide in bacteriophages can be cut through genetic engineering technology. And then insert the gene fragments encoding exogenous polypeptides or proteins between, so that the exogenous polypeptides or protein sequences can form fusion protein molecules with phage capsid proteins to be expressed on the surface of the phage under the guidance of signals. This process is called "display".

10. In general, the preparation and screening of phages expressing specific proteins or polypeptides can be conducted according to the following steps: (1) Prepare a large number of display libraries of phage clones with a specific protein or polypeptide displayed on the surface of each phage. (2) Display libraries are used as mobile phases and target polypeptides or protein molecules as stationary phases to make the two phases contact and interact with each other. (3) Phages that can not specifically bind to target molecule are abandoned. (4) The collected bacteriophages are used for further amplification of E. coli infection, resulting in an enrichment pool as a display library for a new round of elution.

11. Common Methods Part 3 As the time requires, antibody phage display methods and services have been developed and offered by many famous life science companies, including high-quality phage display library construction and custom phage display library screening services that involve specific antibody discovery (e.g. PTM-specific antibody, anti-idiotype antibody, and agonistic antibody) and peptidome discovery.

12. Common Methods Due to the different properties and advantages, the most appropriate phage display system (M13, T4 or T7) is tailored to meet people’s demands. Through DNA manipulation, numerous gene variants can be created and constructed as phage display library. Even generating high-quality libraries with the diversity of 108 becomes possible. Furthermore, novel biopanning strategies receive attention to detect the interaction between displayed protein and those other molecules or isolate specific target binders, and to select high-affinity binders or ligands which are able to recognize the naïve targets. Library Construction and Screening Antibody phage library performs excellent when discovering novel monoclonal antibodies and even the fully human antibody compared with the traditional hybridoma method. Constructing immune antibody libraries and isolating monoclonal antibodies with high specificity and affinity from a comprehensive list of species become operable, which contain but are not restricted to monkey, llama, camel, shark, alligator, mouse, rat, hamster, guinea pig, rabbit, chicken, dog, bovine, goat, sheep, and ferret. Some high-quality pre-made libraries (including human antibody libraries) for our library screening services. Antibody Phage Display Library

13. At present, there still exist some problems in phage display technology, such as limited storage capacity, random folding of long fragment expression structure, which affects biological activity, and obstacles to amino acid modification of host bacteria. Therefore, on the basis of this technology, targeted solutions such as ribosome display technology and yeast protein display system have been further developed. With the continuous development and progress of modern biomedical technology, the system itself will also be constantly prompted and ultimately provide a continuous stream of new ideas and discoveries for the maintenance of human life and health.

14. Company Profile Part 4

15. Creative Biolabs, founded by scientists who are dedicated to the conquering of cancer, is the leading custom service provider that has extensive experience in various antibody production and engineering fields with service portfolio composing mouse and rat monoclonal antibody production adopting hybridoma technology, human, monkey, rabbit, chicken, dog, llama and camel monoclonal antibody production utilizing various antibody library technologies (including phage display, bacterial display and yeast display), which is also professional in conducting in depth antibody humanization and affinity maturation using phage display and DNA mutagenesis approaches. In addition, OEM services for bulk scale antibody manufacturing, including bacterial production of scFv, diabody, tandem scFv, miniantibody and Fab, and mammalian cell expression of minibody, chimeric IgG and IgG, are also available at the most competitive price in the industry. Construction of a custom-service-centered business model is important for optimizing the drug development process, leveraging accessible resources, and forming a team of various background to conduct drug discovery in future.

16. THANK YOU