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 X2 the amount of Ab with Ag T Time 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 T Time f afinity k K=f*p x0 the inchoate value of X1 X0+m the extend of combination

Ab—Ag Ag Key Switchgen

a b

k Ag Ab Key

Variable Descripti

• n

a the ratio

• f

separabili ty between Ab and Ag f afinity k K=f*p b the ratio

• f

separabili ty between Ab and key

X3(0)=X4(0)=X5(0)=0.

‘dx1/dt=a*x5-k*x1*x2’ ‘dx2/dt=-b*x2-k*x1*x2’ ‘dx3/dt=a*x5+b*x2’ ‘dx4/dt=b*x2+k*x1*x2’ ‘dx5/dt=-a*x5+k*x1*x2’,

Variable Description X1 the amount of Ag X2 the amount of Ab with Ag X3 the amount of Ab X4 the amount of key X5 the amount of Ab with key a the ratio of separability between Ab and Ag b the ratio of separability between Ab and key c the inchoate value of X1 d the inchoate value of X2 E the extend of combination t time

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

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

• 1).

Variable Description 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 Ag E the extend of combination d The inchoate value of x1 Variable Description e The inchoate value of x2 t time

• 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