Presentation is loading. Please wait.

Presentation is loading. Please wait.

ANTIBODY ENGINEERING Joud Shafiq. ANTIBODY ENGINEERING ANTIBODIES METHODOLOGIES APPLICATIONS.

Similar presentations


Presentation on theme: "ANTIBODY ENGINEERING Joud Shafiq. ANTIBODY ENGINEERING ANTIBODIES METHODOLOGIES APPLICATIONS."— Presentation transcript:

1 ANTIBODY ENGINEERING Joud Shafiq

2 ANTIBODY ENGINEERING ANTIBODIES METHODOLOGIES APPLICATIONS

3 Heavy chain Light chain 220 a.a ~500 a.a NH2 IMMUNOGLOBULIN

4 Antigen Binding Site Variable Regions Constant Regions Effector Functions

5 LIGHT CHAIN GENES Variable domain CDR3

6 HEAVY CHAIN GENES HUMAN H.C Variable region= V+D+J gene segments

7 ANTIBODY ENGINEERING

8 MONOCLONAL ANTIBODIES Mouse splenocytes Mouse myeloma hybridoma Y Y Y Monoclonal Y Antibodies Y

9 PROBLEMS HAMA Response. Loss of effector functions. Reduce the size of antibody.

10 Production of human Mab. Make mouse Ab more human-like Production of small Ab fragments. SOLUTIONS

11 c c CH1 CH2 CH3 HINGE

12 PROTEASE DIGESTION c c CH1 c c CH3 HINGE F(ab`)2 Fab

13 . HAMA Response. Loss of effector functions. Making the antibody more human-like. (the cutting and pasting) Production of human Mabs. Chimeric AbsHumanized Abs Hybridoma Phage Display Transgenic/SCID Mice

14 CHIMERIC HUMAN/MOUSE VARIABILITY PLOT FR1FR2FR3FR4 CDR1CDR2CDR3 CONSTANT FR1FR2FR3FR4 CDR1CDR2CDR3 Mouse Human Chimeric

15 HumanMouse Chimeric

16 HUMANIZED ANTIBODIES FR1FR2FR3FR4 CDR1CDR2CDR3 CONSTANT FR1FR2FR3FR4 CDR1CDR2CDR3 Mouse Human Humanized

17 HumanMouse Humanized

18 CHIMERIC/HUMANIZED.Reduce the HAMA response..Maintain effector functions..Increase the half life of the antibody..Maintain binding affinity..Easy to construct.

19

20 ANTIBODY ENGINEERING

21 . HAMA Response. Loss of effector functions. Making the antibody more human-like. (the cutting and pasting) Production of human Mabs. Chimeric AbsHumanized Abs Hybridoma Phage Display Transgenic/SCID Mice

22 MONOCLONAL ANTIBODIES Mouse splenocytes Mouse myeloma hybridoma Y Y Y Monoclonal Y Antibodies Y

23 HUMAN HYBRIDOMAS. Unstable hybridomas..Small number of human B-cells in peripheral blood.. Obtaining antigen-reactive human B-cells. Heteromyeloma fusion In vitro immunization

24 MICE SCID Transgenic Reconstituted with human peripheral blood Containing human immunoglobulin genes.

25 PHAGE DISPLAY

26 Reduce the size of Antibody. Production of antibody fragments. Simplifying the expression/production of antibodies.

27 ANTIBODY FRAGMENTS

28 EXPRESSION SYSTEMS Fungal Mammalian cell lines Bacterial

29

30


Download ppt "ANTIBODY ENGINEERING Joud Shafiq. ANTIBODY ENGINEERING ANTIBODIES METHODOLOGIES APPLICATIONS."

Similar presentations


Ads by Google