title: Affinity purification of native glycodelin from amniotic fluid for biological investigations and development of a glycodelin ELISA for clinical studies.
abstract: Glycodelin is a glycoprotein with different oligosaccharides that are responsible for its diverse biological functions in contraception and immunosuppression. Therefore, it is necessary to have access to adequate amounts of glycodelin with retained carbohydrate structure for functional studies because the carbohydrate part can be lacking or be insufficient in recombinant glycodelin from prokaryotic and eukaryotic cell systems.
  
  
  
  Native glycodelin was purified from amniotic fluid by a series of affinity chromatography steps and had many glycosylated forms verified by mass spectrometry. About 7.5 mg glycodelin was obtained from 1.5 L amniotic fluid. No high molecular mass forms of glycodelin were found in amniotic fluid. Aliquots of the purified glycodelin were used as an immunogen in rabbits for antibody production against glycodelin and a calibrator in a highly sensitive glycodelin enzyme-linked immunosorbent assay (ELISA) with a detection limit of about 1 μg/L.
  
  
  
  Native glycodelin was purified from amniotic fluid and used as an immunogen for raising a rabbit antibody against glycodelin and a calibrator in a highly sensitive glycodelin ELISA. We found no high molecular mass forms of glycodelin in amniotic fluid. Aliquots of the purified glycodelin were set aside for functional studies which are in progress.
authors.affiliation: Department of Clinical Biochemistry, Copenhagen University Hospital Hvidovre, Denmark. Electronic address: steen.soerensen@dadlnet.dk.
authors.forename: Steen
authors.initials: S
authors.lastname: Sørensen
authors.affiliation: Department of Clinical Biochemistry, Copenhagen University Hospital Hvidovre, Denmark.
authors.forename: Vibeke
authors.initials: V
authors.lastname: Myrhøj
authors.affiliation: Alphalyse, Odense, Denmark.
authors.forename: Thanh Ha
authors.initials: TH
authors.lastname: Nguyen
authors.affiliation: Alphalyse, Odense, Denmark.
authors.forename: Per
authors.initials: P
authors.lastname: Aaslo
authors.affiliation: Department of Clinical Biochemistry, Copenhagen University Hospital Hvidovre, Denmark.
authors.forename: Young Bae
authors.initials: YB
authors.lastname: Hansen
mesh_terms: D000653:Amniotic Fluid / Q000737:chemistry*; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D000071816:Glycodelin / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D011817:Rabbits
chemical_list: D000906:Antibodies; D000071816:Glycodelin; C045246:PAEP protein, human
keywords: Amniotic fluid; ELISA; Glycodelin; Isolation; Purification