Use of a specific zona pellucida (ZP) protein 3 antiserum as a clinical marker for human ZP integrity and function.
OBJECTIVE
To evaluate binding characteristics of a specific zona pellucida (ZP) protein 3 (ZP3) antiserum to human oocytes in order to determine its usefulness as a clinical marker for human ZP integrity and function and its correlation with IVF outcome.


METHODS
Prospectively designed, blinded, internally controlled study.


METHODS
Tertiary care academic center.


METHODS
Patients undergoing IVF therapy who had either total failed fertilization or partial fertilization were studied.


METHODS
Metaphase II oocytes showing absence of pronuclear formation were salt stored 48 hours after insemination and bisected into matching hemizonae using micromanipulation. One hemizona was incubated with AS ZP3-6 (an antiserum generated against a synthetic ZP3 peptide derived from an amino acid sequence that is highly conserved in the structure of ZP3), whereas the matching hemizona was incubated with AS ZP3-7, an antiserum detecting exclusively mouse ZP3 (internal, negative control). Antibody binding was visualized using the peroxidase-antiperoxidase method and diaminobenzidine as color reagent.


RESULTS
A total of 104 unfertilized oocytes were evaluated. Analysis of variance showed a significant interaction between gamete factor groups (sperm and oocyte) and antiserum factor. Patients with oocyte factor had significantly lower mean staining scores for the AS ZP3-6-treated hemizonae than patients with sperm factor.


CONCLUSIONS
These results demonstrate that anomalies of human ZP3 can be identified with AS ZP3-6 and that these ZP abnormalities correlate with fertilization failure during IVF treatment. Thus, this newly developed biomarker may be of clinical significance in the identification of oocyte defects that are associated with fertilization disorders and may help in the decision-making process in the IVF-assisted fertilization setting.
Oehninger
S
S
Jones Institute for Reproductive Medicine, Eastern Virginia Medical School, Norfolk, USA.
Hinsch
E
E
Pfisterer
S
S
Veeck
L L
LL
Kolm
P
P
Schill
W B
WB
Hodgen
G D
GD
Hinsch
K D
KD
D000328:Adult; D000704:Analysis of Variance; D000818:Animals; D015415:Biomarkers; D004527:Egg Proteins / Q000032:analysis* / Q000276:immunology; D005260:Female; D005307:Fertilization in Vitro; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D011446:Prospective Studies; D011817:Rabbits; D011956:Receptors, Cell Surface; D015044:Zona Pellucida / Q000502:physiology*; D000072216:Zona Pellucida Glycoproteins
D015415:Biomarkers; D004527:Egg Proteins; D007106:Immune Sera; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; C000606517:ZP3 protein, human; D000072216:Zona Pellucida Glycoproteins; C000606521:Zp3 protein, mouse