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2016 ; 11
(9
): e0162973
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English Wikipedia
Anti-Human Platelet Antigen-1a Immunoglobulin G Preparation Intended to Prevent
Fetal and Neonatal Alloimmune Thrombocytopenia
#MMPMID27627660
Weng YJ
; Husebekk A
; Skogen B
; Kjaer M
; Lin LT
; Burnouf T
PLoS One
2016[]; 11
(9
): e0162973
PMID27627660
show ga
Fetal and neonatal alloimmune thrombocytopenia (FNAIT) is a severe disease that
is caused by maternal alloantibodies generated during pregnancy or at delivery as
a result of incompatibility between maternal and fetal human platelet antigens
(HPAs) inherited from the father. Antibody-mediated immune suppression using
anti-HPA-1a immunoglobulins is thought to be able to prevent FNAIT caused by
HPA-1a. A fractionation process to prepare anti-HPA-1a immunoglobulin (Ig) G
(IgG) from human plasma was therefore developed. Anti-HPA-1a plasma was obtained
from volunteer mothers who underwent alloimmunization against HPA-1a during a
previous pregnancy. Plasma was cryoprecipitated and the supernatant treated with
caprylic acid and solvent/detergent (S/D), purified by chromatography,
nanofiltered, concentrated, and sterile-filtered. The anti-HPA-1a immunoglobulin
fraction was characterized for purity and safety. PAK12 and quantitative
monoclonal antibody immobilization of platelet antigen (MAIPA) assays were used
to detect anti-HPA-1a IgG. Hepatitis C virus (HCV) removal during nanofiltration
was assessed by spiking experiments, using cell culture-derived reporter HCV and
luciferase analysis. The caprylic acid treatment precipitated non-Ig proteins
yielding a 90% pure Ig supernatant. S-HyperCel chromatography of the S/D-treated
supernatant followed by HyperCel STAR AX provided high IgG recovery (>80%) and
purity (>99.5%), and efficient IgA and IgM removal. Concentrations of complement
factors C3 and C4 were < 0.5 and < 0.4 mg/dL, respectively. The final IgG could
be nanofiltered on Planova 20N under conditions removing more than 3 log HCV
infectivity to baseline mock infection level, and concentrated to ca. 30 g/L.
Proteolytic activity and thrombin generation were low in the final fraction. The
Pak12 and MAIPA assays showed good recovery of anti-HPA-1a throughout the
process. Clinical-grade HPA-1a IgG can be prepared using a process compliant with
current quality requirements opening perspectives for the prevention of FNAIT.
|Antigens, Human Platelet/immunology/isolation & purification/*therapeutic use
[MESH]
|Chemical Fractionation
[MESH]
|Densitometry
[MESH]
|Electrophoresis, Polyacrylamide Gel
[MESH]
|Hepacivirus
[MESH]
|Humans
[MESH]
|Infant, Newborn/immunology
[MESH]
|Integrin beta3
[MESH]
|Thrombin/metabolism
[MESH]
|Thrombocytopenia, Neonatal Alloimmune/*prevention & control
[MESH]