Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1991-07-03
1996-04-02
Spiegel, Carol A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 792, 435 7021, 43524027, 435968, 436526, 436548, 5303882, G01N 33567
Patent
active
055039812
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a method for the isolation of trophoblast (placental) cells from the blood of a pregnant mammal so as to provide the essential starting material, namely cells derived from the fetus, to enable genetic and/or biochemical information about the fetus to be obtained. In particular, this invention relates to the use of monoclonal antibodies specific for membrane protein markers on mammalian trophoblasts to isolate trophoblast cells from maternal blood. These cells may then be used to obtain fetal genetic and/or biochemical information early in pregnancy. The present invention is particularly relevant for detecting human fetal abnormalities.
Currently, prenatal testing is carried out on fetal cells obtained by either amniocentesis or chorionic villous sampling (CVS). Amniocentesis is normally performed around 16 weeks of gestation. The procedure involves attendance by skilled personnel to insert a needle into the amniotic sac of the fetus and remove between 20-30 ml of amniotic fluid. The amniotic fluid contains fetal cells to allow subsequent tests to be performed. This method of obtaining fetal cells is associated with a risk of inducing a spontaneous abortion. In addition, if the subsequent genetic diagnosis of the fetus reveals an abnormality, the prospect of a mid-trimester pregnancy termination is both psychologically stressful and associated with some physical risk to the mother.
Chorionic villus sampling also requires the involvement of skilled personnel to take a small biopsy from the placenta of an 8-12 week old fetus. Again this procedure has some risk of inducing a spontaneous abortion, although the early diagnosis of any chromosomal abnormality makes the procedure more attractive than amniocentesis. However, the need for skilled personnel and the possibility of inducing spontaneous abortion for both procedures means that current prenatal genetic assessments are made only on pregnant women who are deemed "at risk" of carrying a chromosomally defective fetus.
The method of the present invention provides a simpler procedure which involves obtaining blood from an arm vein of a pregnant mammal such as a pregnant woman and extracting fetal cells which are normally sloughed off from the placenta into the maternal circulation. No specialised expertise is required to obtain this blood sample and this non-invasive isolation of fetal cells negates any risk of inducing a spontaneous abortion. The blood may be taken around the same gestational time as for chorionic villus sampling hence the benefits of early diagnosis are gained.
Although the presence of trophoblast cells in maternal peripheral blood has been the subject of some debate, Goodfellow and Taylor (1982) have reported the extraction of trophoblast cells circulating in peripheral blood during pregnancy by use of differential centrifugation. Covone et al (1984) investigated the possibility of detecting trophoblast cells in the peripheral blood from women at various stages of gestation by the use of monoclonal antibody H315 (Johnson et al, 1981), however in subsequent reports (Pool et al, 1987; Adinolfi et al, 1989), it was suggested that the isolation of H315-positive cells as a source of diagnostic material for antenatal diagnosis of fetal abnormalities is impractical. Recent data suggests that the frequency of fetal cells in the maternal circulation (from 12 weeks to 36 weeks gestation) is less than 1 in 100,000 (Adinolfi et al, 1989; Schwinger et al, 1989).
According to one aspect of the present invention, there is provided a method for the isolation of trophoblast cells from a blood sample of a pregnant mammal which method comprises contacting said blood sample with a binding-effective amount of an antibody specific for villous syncytiotrophoblast and non-villous cytotrophoblast cells for a time and under conditions sufficient for said antibody to bind to target cells and then separating said cells bound by said antibody from said sample.
Another aspect of the present invention is directed to a method for obtaining fetal ge
REFERENCES:
Mueller et al., "Isolation of Fetal Trophoblast Cells from Peripheral Blood of Pregnant Women", The Lancet, vol. 336, pp.197-200, (1990).
Parks et al., "Fetal Cells from Maternal Blood: Their Selection and Prospects for Use in Prenatal Diagnosis", inMethods in Cell Biology,, vol. 26, Academic Press, Inc., pp. 277-295 (1982).
Mueller et al., "Identification of Extra-villous Trophoblast Cells in Human Decidua using an Apparently Unique Murine Monoclonal Antibody to Trophoblast", Hisotchemical Journal, vol. 19, pp. 288-296 (1987).
Nickson et al., "Molecular Cloning and Expression of Human Trophoblast Antigen FD0161G and Its Identification as 3.beta.-hydroxy-5-ene Steroid Dehydrogenase", J. Reprod. Fert., vol. 93, pp. 149-156 (1991).
Molday et al., Separation of Cells Labeled with Immunospecific Iron Dextran Microsheres Using High Gradient Magnetic Chromatography, vol. 170, No. 2, pp. 323-238 (1984).
Pourfarzaneh et al., "The Use of Magnetizable Particles in Solid Phase Immunoassay", in Methods of Biochemical Analysis, D. Glick, ed., John Wiley, New York, vol. 28, pp. 267-295, (1981).
Mueller, U. W., Histochemical Journal 19, 288-296 (1987).
Sen-Majumdar, A., Biochemistry 25, 634-640 (1986).
Chemical Abstracts 103:192486c.
Chemical Abstracts 104:82124w.
Chemical Abstracts 102:44039y.
Butterworth et al--Chem. Abst. vol. 103 (1985) p. 192,486c.
Kawata et al--Chem. Abst. vol. 101 (1984) p. 149,452k.
Biological Abstracts, vol. 73, 47702 (1982).
Biological Abstracts, vol. 84, 71902 (1987).
Biological Abstracts, vol. 87, 35303 (1989).
Biological Abstracts, vol. 87, 35304 (1989).
Chemical Abstracts, 105:40798j (1986).
Hawes Catherine S.
Mueller Utz W.
Flinders Technologies Pty, Ltd.
Spiegel Carol A.
Wolski Susan C.
LandOfFree
Isolation of fetal cells from maternal blood to enable prenatal does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Isolation of fetal cells from maternal blood to enable prenatal , we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Isolation of fetal cells from maternal blood to enable prenatal will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2015778