Generation and characterization of a polyclonal antipeptide antibody to human glycodelin.
OBJECTIVE
To develop and characterize an antiglycodelin antibody using a 15-amino acid synthetic peptide as antigen, derived from the sequence of human glycodelin.


METHODS
We have developed a chicken antiglycodelin-derived peptide antibody and have characterized the antibody with the use of endometrial and ovarian cell lines. The antibody was also tested for its ability to detect glycodelin by ELISA assay, immunocytochemistry, and by Western blot.


METHODS
Various cell lines, cell culture medium, and amniotic fluid were used in the experiments.


METHODS
Amniotic fluid was collected from pregnant patients in their first trimester of pregnancy.


METHODS
No intervention.


METHODS
Detection of glycodelin.


RESULTS
The cell lines RL95-2 (human endometrial carcinoma cells), OVCAR-3 (human ovarian adenocarcinoma cells), HeLa (human cervical epitheloid carcinoma cells), and EM42-D (human endometrial epithelial cells) reacted with the antibody, indicating the presence of glycodelin. A specific 45-kd protein representing glycodelin was detected by Western blot in the amniotic fluid.


CONCLUSIONS
Antipeptide antibodies can be successfully used to detect and quantify the presence of glycodelin in cells and fluids.
Poddar
A S
AS
Department of Gynecology and Obstetrics, Emory University, Atlanta, Georgia 30322, USA.
Kim
J G
JG
Gill
K P
KP
Bates
B N
BN
Santanam
N
N
Rock
J A
JA
Murphy
A A
AA
Parthasarathy
S
S
D000230:Adenocarcinoma / Q000737:chemistry; D000653:Amniotic Fluid / Q000737:chemistry; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D002645:Chickens / Q000276:immunology; D016889:Endometrial Neoplasms / Q000737:chemistry; D004717:Endometrium / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000737:chemistry; D005260:Female; D000071816:Glycodelin; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology*; D006367:HeLa Cells / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D010051:Ovarian Neoplasms / Q000737:chemistry; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000032:analysis / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000071816:Glycodelin; D006023:Glycoproteins; C045246:PAEP protein, human; D011257:Pregnancy Proteins