Observations on Marketed Alpha-1-Proteinase Inhibitor Products Ewa Marszal, PhD FDA/CBER
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Observations on Marketed Alpha-1-Proteinase Inhibitor Products
Ewa Marszal, PhD
FDA/CBER
BPAC, November 2005
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Presentation Contents
Differences between 1-PI products
Consistency of manufacturing
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Differences between 1-PI products
Contaminating plasma proteins
Inactive 1-PI (e.g., polymer/latent form)
Modifications of the primary structure
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Protein impurities(SDS-PAGE, reducing)
Prolastin Aralast Zemaira
albumin →1-PI →
Specific activity: ≥ 0.35 ≥ 0.55 ≥ 0.7mg functional 1-PI/mg total protein
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-4 0 0 0 0 0
-2 0 0 0 0 0
0
2 0 0 0 0 0
4 0 0 0 0 0
6 0 0 0 0 0
8 0 0 0 0 0
1 E +0 6
1 E +0 6
0 5 1 0 1 5 2 0 2 5 3 0 3 5
Presence of high molecular weight species(SE-HPLC, absorbance@215 nm)
M%
95.4
89.8
88.1
Zemaira
Aralast
ProlastinM
1-PI + albumin)
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Isoelectric focusing (IEF)
cathode (-)
anode (+)
pH gradient
higher pH
lower pH
zero net charge
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Heterogeneity of 1-PI in plasma by IEF(charge heterogeneity)
3 polysaccharide chains (sialic acid)
N-terminal pentapeptide EDPNG
Polymorphism, e.g., E342K
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EDPNG N N N
EDPNG N N NM6
M4
M2
Dominant 1-PI isoforms in plasma (charge heterogeneity)
EDPNG N N N
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Heterogeneity of 1-PI products by IEF P
lasm
a
Ara
last
Zem
aira
Pro
last
in
Pla
sma
Ara
last
Zem
aira
Pro
last
in
desialylated
M6 → M4 → M2 →
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Consistency of manufacturing by IEF
Analysis of patients’ plasma from Aralast pivotal clinical trial
Analysis of current and historical lots of 1-PI products
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Conclusions
1-PI products contain: different levels of contaminating plasma proteins different amounts of inactive 1-PI species different levels of primary structure modifications
1-PI in each product differs somewhat from 1-PI in plasma (IEF, polymer)
Manufacturing of each 1-PI product appears to be consistent over time by IEF
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Consistency of manufacturing
Bayer Baxter Behring
Desialylated
Non-desialylated