Structural & functional characterisation of antibody therapeutics

71 Pages·2010·3.78 MB·English

Structural & functional characterisation of antibody
therapeutics
Digitalis
Digitalis
Roy Jefferis
School of Immunity & Infection, University of Birmingham UK
CASSS – Practical Applications of Mass Spectrometry.
Marina del Rey, September 2010
Market size: $22 billion in 2007
Projected for 2010: $30 billion
28 licensed; ~ 500 in development
80 in clinical trials; 200 companies
Challenge: To reduce:
“Cost of Goods” (CoG)
“Cost of Treatment” (CoT)
Natural & recombinant antibodies protect due to:
Neutralisation/blocking of infectious agents and their products:
e.g. virus, toxins, venoms cytokines, e.g. TNFα
Infliximab, Adalimumab, Golimumab, Cimzia(Fab-Peg)
Formation of immune complexes and activation of killing
mechanisms e.g. bacteria cancer cells
Rituximab, Herceptin, Cetuximab
Agonist or antagonist activity (autoimmunity!)
Panitumumab, Cetuximab
Structural & functional characterisation of antibody
therapeutics
Antibody classes, isotypes and allotypes
Antibody effector activities
IgG-Fc glycoforms
IgG-Fab glycoforms
Biosimilars & Antibody-drug conjugates
Human immunoglobulin classes & subclasses
The result of gene duplication, mutation and selection
IgM IgG IgD IgE IgA
IgG1, IgG2, IgG3, IgG4; IgA1, IgA2
The humoral immune response is “orchestrated” to provide
optimal protection to a given “insult”
Four chain structure of the antibody molecule
Hinge region, susceptible to enzymatic cleavage
V C
L L
V C 1 C 2 C 3
H H H H
Fab: antigen binding Fc: effector activation
Oligosaccharide
Interchain disulphide bridges
Domain structure of IgG
Antigen
Antigen
binding
binding
Fab
Fab
oligosaccharide
Fc
Effector functions
Ludger
IIggGG22
IIggGG11
~ 25 %
~ 60 %
SpA/SpG binding
SpA/SpG binding
IIggGG44
IIggGG33
~ 5 %
~ 10 %
SpA/SpG binding
SpG binding
IgG subclass of licensed antibody therapeutics
Rituxan IgG1κ
Golimumab IgG1κ
Zenopax IgG1κ
Xolair IgG1κ
Herceptin IgG1κ
Raptiva IgG1κ
Remicade IgG1κ
Erbitux IgG1κ
Simulect IgG1κ
Avastin IgG1κ
Synagis IgG1κ
Mylotarg IgG4κ
Campath IgG1κ
Tysabri IgG4κ
Humira IgG1κ
Mylotarg IgG4κ
Tocilizumab IgG1κ
Vectibix IgG2κ
Biacore binding signals (RU) & dissociation constants (Kdiss) for
anti-IgG-Fc: R10Z8E9 with human & non-human primate sera
Sample (serum) RU KDiss (M)
Human 1274 1.77 ×10−10
Cynomolgus 3 No binding
Baboon 0 No binding
Marmoset 5 No binding
Rhesus macaque −3 No binding
Chimpanzee 1077 2.21 ×10−10
Stubenrauch K, et al., J Pharm Biomed Anal. 49:1003-1008 (2009)