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 Antigen (affinity stronger) Mutated Antigenic Determinant (affinity weaker) both tend to combine with Antibody Finally, the stronger one win!

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

Background Knowledge What inspired us?

ADCs & Inspiration

ADCs & Inspiration

ADCs • Benefits? • Flaws? • Solutions?

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

With Switchgen: Enhanced CAR-T

Design How to construct it?

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

Experiment Based on FRET

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

Modeling Further on how to demonstrate it

• 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

• 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

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

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

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

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

• 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.

Human Practice • Our own We-Chat Public Platform

Human Practice • Investigation 1

Human Practice Investigation 2

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

Human Practice • Meet up • Communication with Tito Jankowski

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

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.

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

Attribution • Instructor: Peilin Li • Advisors: Boxiang Wang, Yuying Zhang • Sponsors

Thank you! Shenzhen_SFLS