Download CSA Biopharm - Moodle Lille 2

Survey
yes no Was this document useful for you?
   Thank you for your participation!

* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project

Document related concepts

Immunocontraception wikipedia , lookup

Immunoprecipitation wikipedia , lookup

Adoptive cell transfer wikipedia , lookup

Duffy antigen system wikipedia , lookup

Antimicrobial peptides wikipedia , lookup

Antibody wikipedia , lookup

Major histocompatibility complex wikipedia , lookup

Gluten immunochemistry wikipedia , lookup

DNA vaccination wikipedia , lookup

Immunomics wikipedia , lookup

Cancer immunotherapy wikipedia , lookup

Immunosuppressive drug wikipedia , lookup

ELISA wikipedia , lookup

Monoclonal antibody wikipedia , lookup

Molecular mimicry wikipedia , lookup

Polyclonal B cell response wikipedia , lookup

Human leukocyte antigen wikipedia , lookup

Transcript
Séminaire de Communication Scientifique
Faculté de Pharmacie de Lille
20 février 2014
DESPRES Chloé
NOURRY Sandra
OLIVIER Agathe
IMMUNOGENICITY
OF BIOLOGICS
Prediction tools & Risks assessment
CSA Biopharm
1
CSA Biopharm
CEO
Chief Medical
Officer
Chief Scientic
Officer

Goal : to develop the pipeline in biotechnologies

Just discovered a new mouse mAb for Alzheimer’s
disease
2
CSA Biopharm
CSA Biopharm
CEO
Chief Medical
Officer
Chief Scientic
Officer
We want to be sure there will
be
no
immunogenicity
problems that could stop the
development during the clinical
trials and later.
3
CSA Biopharm
Humanization
4
CSA Biopharm
Humanization : HOW ?
Goal :
Human antibody (80%) :
reduce immunogenicity
•
•
•
AHAM : anticorps humains anti-anticorps murins
AHAC : anti-corps humains anti-anticorps
chimériques
AHAH : anticorps humains anti-anticorps
humanisés
5
CSA Biopharm
Examples
Antibody name
Company
Type
Target
Indication(s)
Reported
immunogenicity
Infliximab
(REMICADE)
Centocor
(J&J)
Chimeric
TNFα
RA/Crohn
10-15%
Panitumumab
(VECTIBIX)
Amgen
Human
EGFR
Colorectal
Cancer
4,6%
Ustekinumab
(STELARA)
Centocor
(J&J)
Human
IL12-IL23
Plaque
Psoriasis
3-5%
TNFα
RA/Crohn/
Ankylosing
spondylitis/
plaque
psoriasis…
2.6%–26%
Adalimumab
(HUMIRA)
Abbott
Human
6
Matthew P Baker et Al. Immunogenicity of protein therapeutics:The key causes, consequences and challenges. Self/Nonself 1:4, 314-322;
October/November/December 2010 © 2010 Landes Bioscience
Humanization : HOW ?

Goal :
-
Murines CDR (20%) : responsible for the desired binding properties

Risk :
-
Decrease of affinity and activity.
-
Creation of new epitope T at junctions and induced immunogenicity and side effects.
7
CSA Biopharm
Humanization : illustrating

Not just insert murines CDR
8
CSA Biopharm
Examples
Antibody name
Company
Type
Target
Indication(s)
Reported
immunogenicity
Infliximab
(REMICADE)
Centocor
(J&J)
Chimeric
TNFα
RA/Crohn
10-15%
Panitumumab
(VECTIBIX)
Amgen
Human
EGFR
Colorectal
Cancer
4,6%
Ustekinumab
(STELARA)
Centocor
(J&J)
Human
IL12-IL23
Plaque
Psoriasis
3-5%
TNFα
RA/Crohn/
Ankylosing
spondylitis/
plaque
psoriasis…
2.6%–26%
Adalimumab
(HUMIRA)
Abbott
Human
9
Matthew P Baker et Al. Immunogenicity of protein therapeutics:The key causes, consequences and challenges. Self/Nonself 1:4, 314-322;
October/November/December 2010 © 2010 Landes Bioscience
Immune Response to Therapeutic Protein :
Why is it important ?

Development of Anti-Drug Antibody (ADA)

Affect both efficacy and safety of the product
1.
Result in reduced efficacy and sometimes of a complete lack of a clinical response
2.
Result in adverse events (benign to life-threatening)

Hypersensibility…
10
CSA Biopharm
And for the regulatory
agencies ?

Widespread use therapeutic proteins has demonstrated that
nearly all biologicals can elicit antibody responses
FDA & EMA demand :
No production of ADA
causing adverse effect

Animal models are not predictive for immunogenicity :
development of anti human protein antibody
11
CSA Biopharm
Immunogenicity :
Predictive Tools
12
CSA Biopharm
But before…
… The Immunogenicity
mechanism
13
CSA Biopharm
How does it work ?
CD/LcB
http://www.youtube.com/watch?v=Mn9_sgUnemk
14
CSA Biopharm
Immunogenicity tests
16
CSA Biopharm
T Cell Epitope Antigenicity
Profiling

In silico assays

In vitro assays

Class II HLA binding assays

Antigen presentation assays

T cell proliferation assays

Cytokines release assays
17
CSA Biopharm
Immunogenicity tests
In silico
18
CSA Biopharm
In silico binding prediction
19
CSA Biopharm
In silico binding prediction
Macromolecular Modeling Blog ™
Zavala-Ruiz Z et al. PNAS 2004;101:13279-13284
20
CSA Biopharm
In silico binding prediction
Goal : prediction of MHC II/ peptide interaction
Ab is virtually splited  peptides
Peptides created are shifted by 1 amino acid
AEYILPMKFDVSKLGFQ
AEYILPMKFDVS
EYILPMKFDVSK
YILPMKFDVSKL
ILPMKFDVSKLG
LPMKFDVSKLGF
PMKFDVSKLGFQ
21
CSA Biopharm
In silico – Results
L. Moise et Al. Effect of HLA DR epitope de-immunization of Factor VIII in vitro and in vivo.ClinicalImmunology (2012)
142, 320–331
22
CSA Biopharm
Deimmunization
YILPMKFDVSKL
YGLPMKFDVSGL

Identify and eliminate T-cell epitopes from the variable
region sequences of antibodies and proteins.
23
CSA Biopharm
De-immunization : it is
working!
24
CSA Biopharm L. Moise et Al. Effect of HLA DR epitope de-immunization of Factor VIII in vitro and in vivo.ClinicalImmunology (2012)
In Silico Strengths and Limitations
High throughput
No proof of TCR interaction
Low cost
Consequences of this interaction?
(production of Ab or T rég, affinity…)
Reduce downstream in vitro testing
No proof of real interaction
Opportunity in early development to
modify a protein to decrease its
immunogenic potential
25
Immunogenicity tests
In vitro
26
Class II HLA-peptide Binding Assay
27
Class II HLA-peptide Binding Assay
Goal :
Determines the ability of each candidate peptide to bind class II HLA alleles and to
stabilize the HLA-peptide complex
28
HLA Binding Assay Strengths and
Limitations
Straight forward & Easier
No proof of TCR interaction
Improve accuracy of immunogenicity
predictions before using biological
assay
Artificial processing
29
Antigen Presentation Assays
30
How does it work ?
Culture of the protein of interest with
DCs.DCs take up the protein and process
it
HLA molecules present epitopes
from the protein on the DC surface.
Cells are lyzedand HLApeptide molecules are
then recovered in an
immune affinity step
and analyzed by sequencing mass
spectrometry.
Peptides are recovered from
the HLA
CSABiopharm
Peptides identified by
mass spectrometry are
subjected to analysis to
identify true positive
peptides with high
32
confidence.
Antigen Presentation Assay
Strengths and limitations
Inform about natural antigen
processing.
No proof of TCR interaction and T cell
proliferation
Assessment of MHC peptide binding
Rapid way to identify sequences of key
relevance for the immunogenicity.
36
T-Cell Activation
37
T-cell proliferation –
Fluorescent marker dilution
Using whole protein or synthesized peptide
Flow Cytometry
39
T-cell proliferation –
Fluorescent marker dilution
40
Cytokine Release Assay
PRINCIPLE :
ELISA Sandwich
41
X8
X9
X4
X5
X6
X7
PBS
SEB
X1
X2
X3
Results
42
T-cell Activation Assay
Strengths and limitations
Brings together all the partners of
immunogenicity
Risk of variability
Difficulties of reproductibility
Incorporate Ag natural processing and
presentation pathways
Overestimation of the frequency of
activated T cells
Number of individual blood (PBMC) is
quite large (> 40)
 HLA diversity of a patient population
43
TESTS - summary


In silico

Algorithm to screen for potential T-cell epitopes

Identify linear motifs of 9-10 amino-acids that bind to MHC Class II
molecules

Fast, extensive databases exist

Tend to over-predict potential for immune response relative to in vitromethods

Typically used as part of lead selection/optimization
In vitro

HLA binding assays

Antigen Presentation Assay

T cells assays


Peptides or proteins
Typically used as part of lead selection/optimization
44
Decision Tree
+ affinity assays
 efficacy
Humanized mAB
De-immunization beginning
modifications
immunogenic sequences ? (In silico tests)
« in silico » High immunogenicity « in silico » Low immunogenicity
Overlapping peptides production
HLA-binding test
Ag Presentation test
High immunogenicity
T-cell activation test
Low immunogenicity
mAB candidates
Choice of the best mAB (Safety/Efficacy)
Olinoudezumab
Mouse model
46
Thank you for
your attention !
Question ?
47