When the light went out.. All you can think of Switch! Switchgen - PowerPoint PPT Presentation

When the light went out.. All you can think of Switch! Switchgen Shenzhen_SFLS How does it work? How to construct it? How to apply it? How to demonstrate it? Further on How does it work? Basic Mechanism: Original

1. When the light went out.. All you can think of…

2. Switch!

3. Switchgen Shenzhen_SFLS

4. • How does it work? • How to construct it? • How to apply it? • How to demonstrate it? • Further on

5. How does it work? Basic Mechanism: Original Antigen (affinity stronger) Mutated Antigenic Determinant (affinity weaker) both tend to combine with Antibody Finally, the stronger one win!

6. How does it work? • Key – Antigenic Determinate • Circuit – Competitive Binding • Appliance – Effector Protein

7. Background Knowledge What inspired us?

10. ADCs • Benefits? • Flaws? • Solutions?

11. CAR-T • CAR: chimeric antigen receptor • T: T lymphocyte • scFv: single chain fragment variable

12. With Switchgen: Enhanced CAR-T

13. Design How to construct it?

14. How to construct it? • Goal: Proper interactions – Structural Modeling – Protein Fusing – Confirmatory Experiment

15. Experiment Based on FRET

16. Oh no! What should we do? • Key word: practical • DNA construction: direct synthesis (no more PCR!!!) • Yeast or No?

17. Modeling Further on how to demonstrate it

18. • Ab with wAg + one Ag coming into one Ab with Ag and one wAg is irreversible. • dx1/dt=dx2/dt=-k*x1*x2’ • ’k=pf’ • ‘x2=x1+m Variable Description X1 the amount of Ag the amount of Ab with Ag X2 Time T f afinity k K=f*p x0 the inchoate value of X1 X0+m the extend of combination

19. • x1= m/(exp(m*(log((m + c)/c)/m + f*p*t)) - 1) • ‘E=(x0-x1)/x1’ which is simplified as • ‘E=1- m/((exp(m*(log((m + c)/c)/m + f*p*t)) - 1)*c) Variable Description X1 the amount of Ag X2 the amount of Ab with Ag Time T f afinity k K=f*p x0 the inchoate value of X1 X0+m the extend of combination

20. Variable Descripti on a the ratio Key Ag of separabili ty a Ab—Ag between Ab and Ag f afinity k Ab b k K=f*p Switchgen Ag Key the ratio b of separabili ty between Ab and key

21. Variable Description the amount of Ag X1 X2 the amount of Ab with Ag the amount of Ab X3 the amount of key ‘dx1/dt=a*x5-k*x1*x2’ X4 X5 the amount of Ab with key ‘dx2/dt=-b*x2-k*x1*x2’ the ratio of separability between Ab a ‘dx3/dt=a*x5+b*x2’ and Ag ‘dx4/dt=b*x2+k*x1*x2’ the ratio of separability between Ab b ‘dx5/dt=-a*x5+k*x1*x2’, and key c the inchoate value of X1 X3(0)=X4(0)=X5(0)=0. d the inchoate value of X2 E the extend of combination time t

22. Description Variable the amount of Ag X1 the ratio of separability between Ab and Ag a the ratio of separability between Ab and key b c the inchoate value of X1 d the inchoate value of X2 E the extend of combination time t

23. dx1/dt=a*x1-b*x1*x2 ……1 dx2/dt=c*x2-b*x1*x2 ……2 'x1(0)=d''x2(0)=e' x1 =-(d - (t*(c*(e + a*c*t) - a*c))/b)/(a*c*t^2 Variable Description - 1). X1 the amount of Ag X2 the amount of Ab with key b the possibility of the occurrence of the reaction c the ratio of the proliferation of Ab with key a the ratio of the proliferation of Variable Description Ag e The inchoate value of x2 E the extend of combination t time d The inchoate value of x1

24. • The Lotka–Volterra equations , also known as the predator–prey equations , are a pair of first-order, non-linear, differential equations frequently used to describe the dynamics of biological systems in which two species interact, one as a predator and the other as prey.

25. Human Practice • Our own We-Chat Public Platform

26. Human Practice • Investigation 1

27. Human Practice Investigation 2

28. Human Practice • Collaborations - SZMS 15 Shenzhen - South China University of Technology

29. Human Practice • Meet up • Communication with Tito Jankowski

30. Referrence • [1] Aaron C. Anselmo, Samir Mitragotri, An Overview of Clinical and Commercial Impact of Drug Delivery Systems, Journal of Controlled Release (2014), doi: 10.1016/j.jconrel.2014.03.053 • [2] James T. Wu, c-erbB2 oncoprotein and its soluble ectodomain: a new potential tumor marker for prognosis early detection and monitoring patients undergoing Herceptin treatment, Clinica Chimica Acta (2002) • [3] Zuhaida Asra Ahmad, Swee Keong Yeap,Abdul Manaf Ali, Wan Yong Ho, Noorjahan Banu Mohamed Alitheen, and Muhajir Hamid, scFv Antibody: Principles and Clinical Application, Clinical and Developmental Immunology, Volume 2012, Article ID 980250, 15 pages, doi:10.1155/2012/980250

31. Referrence • [4] SWISS-MODEL Workspace: • Marco Biasini, Stefan Bienert, Andrew Waterhouse, Konstantin Arnold, Gabriel Studer, Tobias Schmidt, Florian Kiefer, Tiziano Gallo Cassarino, Martino Bertoni, Lorenza Bordoli, Torsten Schwede (2014). SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information Nucleic Acids Research 2014 (1 July 2014) 42 (W1): W252-W258; • Arnold K, Bordoli L, Kopp J, and Schwede T (2006). The SWISS-MODEL Workspace: A web- based environment for protein structure homology modelling. Bioinformatics.,22,195-201. • Bordoli, L., Kiefer, F., Arnold, K., Benkert, P., Battey, J. and Schwede, T. (2009). Protein structure homology modelling using SWISS-MODEL Workspace. Nature Protocols, 4,1. SWISS-MODEL Repository: • Kiefer F, Arnold K, Künzli M, Bordoli L, Schwede T (2009). The SWISS-MODEL Repository and • associated resources. Nucleic Acids Res. 37, D387-D392. • Kopp J, and Schwede T (2006). The SWISS-MODEL Repository: new features and functionalities. Nucleic Acids Res.,34, D315-D318. • SWISS-MODEL and Swiss PdbViewer • Guex, N., Peitsch, M.C. Schwede, T. (2009). Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: A historical perspective. Electrophoresis, 30(S1), S162-S173.

32. Attribution • Theory Design Group: Yuhe Wu, Yi Zhang, Zhaoheng Li, Qixin Lin • Experiment Group: Jiazheng Xing, Jiafu Li, Zhulin Chen, Fanghui He • Modeling Group: Zheshen Gong, Yan Xu • Publicity Group: Wenjing Jiang • Wiki Making: Junhao Cui