title: A comparison of the choice of monoclonal antibodies for recovery of fetal cells from maternal blood using FACS for noninvasive prenatal diagnosis of hemoglobinopathies.
abstract: BACKGROUND
  Isolation of adequate numbers of fetal cells circulating in the maternal circulation is the major hurdle in developing noninvasive prenatal diagnostic procedures. We used flow cytometry and a combination of different monoclonal antibodies to compare the yield and purity of the fetal nucleated red blood cells at different periods of gestation.
  
  
  METHODS
  Using a Percoll discontinuous gradient, the fetal nucleated erythrocytes were enriched from 7 ml maternal blood. In 100 samples, the enriched cells were stained with CD45, anti-fetal hemoglobin, and glycophorin A antibodies and in 30 samples they were stained with CD45, anti-fetal hemoglobin, and CD71 and then sorted and used for fetal diagnosis of hemoglobinopathies.
  
  
  RESULTS
  Using the first set of antibodies, although we were able to obtain a higher percentage of fetal nucleated red cells (0.07% +/- 0.2%) as compared to the second set which yielded comparatively smaller numbers (0.025% +/- 0.03%), there was some compromise in purity.
  
  
  CONCLUSIONS
  Using CD45, anti-fetal hemoglobin and CD71 would be preferred as minimizing maternal contamination is more important than yield for prenatal diagnosis.
authors.affiliation: National Institute of Immunohaematology, K.E.M. Hospital Campus, Parel, Mumbai, India.
authors.forename: Edna
authors.initials: E
authors.lastname: D'Souza
authors.forename: Kanjaksha
authors.initials: K
authors.lastname: Ghosh
authors.forename: Roshan
authors.initials: R
authors.lastname: Colah
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D015703:Antigens, CD / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D004912:Erythrocytes / Q000276:immunology* / Q000473:pathology; D005260:Female; D005315:Fetal Diseases / Q000175:diagnosis* / Q000276:immunology; D005319:Fetal Hemoglobin / Q000276:immunology; D005333:Fetus / Q000166:cytology*; D005434:Flow Cytometry; D006021:Glycophorins / Q000276:immunology; D006453:Hemoglobinopathies / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D017493:Leukocyte Common Antigens / Q000276:immunology; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011262:Pregnancy Trimester, Second; D011296:Prenatal Diagnosis / Q000379:methods*; D011990:Receptors, Transferrin / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C068126:CD71 antigen; D006021:Glycophorins; D011990:Receptors, Transferrin; D005319:Fetal Hemoglobin; D017493:Leukocyte Common Antigens