Development and characterization of monoclonal antibodies for the immunohistochemical detection of glycodelin A in decidual, endometrial and gynaecological tumour tissues.
OBJECTIVE
Glycodelin is a glycoprotein with a molecular weight of 28 kDa. Unusual LacdiNAc structures have been identified on glycodelin A, isolated from amniotic fluid. Three major functions of this glycoprotein have been identified. Glycodelin is an immunosuppressive molecule, a marker of morphological differentiation, and a contraceptive. Because no monoclonal antibodies for glycodelin A are commercially available, our aim was to develop and characterize three monoclonal antibodies against this glycoprotein.


RESULTS
Glycodelin A was purified from amniotic fluid by three chromatographic steps and its purity was checked by SDS-PAGE. Antibodies were generated from immunized BALB/c mice. Three IgG1 monoclonal antibodies detecting glycodelin A were cloned. All three antibodies recognized carbohydrate structures of glycodelin A and did not cross-react with glycodelin S. They are applicable to immunohistochemistry (frozen and paraffin sections), ELISA and Western blots.


CONCLUSIONS
The new antibodies can be used for the detection of glycodelin A in frozen and paraffin-embedded decidual and endometrial tissue. One antibody (A87-B/D2) can be used for the detection of glycodelin in endometrial and ovarian tumour tissues. Because glycodelin A is a major secretory endometrial product during the luteal phase, in early pregnancy and in gynaecological tumours, the new antibodies are, potentially, valuable tools for the study of endometrial development and tumour progression.
Jeschke
U
U
Department of Obstetrics and Gynaecology, Ludwig-Maximilians-University of Munich, Munich, Germany. udo.jeschke@med.uni-muenchen.de
Kuhn
C
C
Mylonas
I
I
Schulze
S
S
Friese
K
K
Mayr
D
D
Speer
R
R
Briese
V
V
Richter
D-U
DU
Haase
M
M
Karsten
U
U
D000653:Amniotic Fluid / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D002241:Carbohydrates / Q000276:immunology; D003656:Decidua / Q000737:chemistry*; D016889:Endometrial Neoplasms / Q000378:metabolism*; D004717:Endometrium / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000737:chemistry; D005260:Female; D000071816:Glycodelin; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D010051:Ovarian Neoplasms / Q000378:metabolism; D011257:Pregnancy Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D000071816:Glycodelin; D006023:Glycoproteins; C045246:PAEP protein, human; D011257:Pregnancy Proteins