Development, characterization, and use of monoclonal antibodies made to antigens expressed on the surface of fetal nucleated red blood cells

Francisco V. Alvarez, Jitka Olander, Dan Crimmins, Belén Prieto, Ana Paz, Rebeca Alonso, Sharon Porter, Jay Hess, Robert D. Crist, Yvonne Landt, Jack H. Ladenson

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background: Current methods for obtaining fetal cells for prenatal diagnosis are invasive and carry a small (0.5-1.0%) but definite risk of miscarriage. An attractive alternative would be isolation of fetal cells from peripheral maternal blood using antibodies with high specificity and avidity. Methods: To generate antibodies, we purified nucleated red blood cells (NRBCs) from fetal livers and used them as the immunogen to generate monoclonal antibodies (mAbs) directed against surface antigens. Results: The four antibodies recognized at least two conformationally sensitive epitopes of the transferrin receptor. Isolation of NRBCs from 252 maternal blood samples using these antibodies in magnetic activated cell sorting after an initial density gradient centrifugation yielded 0-419 NRBCs per 25 mL of maternal blood. One antibody, 2B7.4, not only isolated the highest number of NRBCs (>10 in 90% of the samples) but also isolated these NRBCs in 78 consecutive maternal samples. Conclusion: Antibody 2B7.4 shows promise for the isolation of NRBCs from maternal blood and should allow studies concerning the source of these cells, fetal vs maternal, and the factors controlling their prevalence.

Original languageEnglish (US)
Pages (from-to)1614-1620
Number of pages7
JournalClinical Chemistry
Volume45
Issue number9
StatePublished - 1999
Externally publishedYes

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Blood
Erythrocytes
Monoclonal Antibodies
Cells
Mothers
Antigens
Antibodies
Transferrin Receptors
Density Gradient Centrifugation
Cell Separation
Spontaneous Abortion
Surface Antigens
Prenatal Diagnosis
Epitopes
Centrifugation
Sorting
Liver

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Alvarez, F. V., Olander, J., Crimmins, D., Prieto, B., Paz, A., Alonso, R., ... Ladenson, J. H. (1999). Development, characterization, and use of monoclonal antibodies made to antigens expressed on the surface of fetal nucleated red blood cells. Clinical Chemistry, 45(9), 1614-1620.

Development, characterization, and use of monoclonal antibodies made to antigens expressed on the surface of fetal nucleated red blood cells. / Alvarez, Francisco V.; Olander, Jitka; Crimmins, Dan; Prieto, Belén; Paz, Ana; Alonso, Rebeca; Porter, Sharon; Hess, Jay; Crist, Robert D.; Landt, Yvonne; Ladenson, Jack H.

In: Clinical Chemistry, Vol. 45, No. 9, 1999, p. 1614-1620.

Research output: Contribution to journalArticle

Alvarez, FV, Olander, J, Crimmins, D, Prieto, B, Paz, A, Alonso, R, Porter, S, Hess, J, Crist, RD, Landt, Y & Ladenson, JH 1999, 'Development, characterization, and use of monoclonal antibodies made to antigens expressed on the surface of fetal nucleated red blood cells', Clinical Chemistry, vol. 45, no. 9, pp. 1614-1620.
Alvarez, Francisco V. ; Olander, Jitka ; Crimmins, Dan ; Prieto, Belén ; Paz, Ana ; Alonso, Rebeca ; Porter, Sharon ; Hess, Jay ; Crist, Robert D. ; Landt, Yvonne ; Ladenson, Jack H. / Development, characterization, and use of monoclonal antibodies made to antigens expressed on the surface of fetal nucleated red blood cells. In: Clinical Chemistry. 1999 ; Vol. 45, No. 9. pp. 1614-1620.
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AU - Alvarez, Francisco V.

AU - Olander, Jitka

AU - Crimmins, Dan

AU - Prieto, Belén

AU - Paz, Ana

AU - Alonso, Rebeca

AU - Porter, Sharon

AU - Hess, Jay

AU - Crist, Robert D.

AU - Landt, Yvonne

AU - Ladenson, Jack H.

PY - 1999

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N2 - Background: Current methods for obtaining fetal cells for prenatal diagnosis are invasive and carry a small (0.5-1.0%) but definite risk of miscarriage. An attractive alternative would be isolation of fetal cells from peripheral maternal blood using antibodies with high specificity and avidity. Methods: To generate antibodies, we purified nucleated red blood cells (NRBCs) from fetal livers and used them as the immunogen to generate monoclonal antibodies (mAbs) directed against surface antigens. Results: The four antibodies recognized at least two conformationally sensitive epitopes of the transferrin receptor. Isolation of NRBCs from 252 maternal blood samples using these antibodies in magnetic activated cell sorting after an initial density gradient centrifugation yielded 0-419 NRBCs per 25 mL of maternal blood. One antibody, 2B7.4, not only isolated the highest number of NRBCs (>10 in 90% of the samples) but also isolated these NRBCs in 78 consecutive maternal samples. Conclusion: Antibody 2B7.4 shows promise for the isolation of NRBCs from maternal blood and should allow studies concerning the source of these cells, fetal vs maternal, and the factors controlling their prevalence.

AB - Background: Current methods for obtaining fetal cells for prenatal diagnosis are invasive and carry a small (0.5-1.0%) but definite risk of miscarriage. An attractive alternative would be isolation of fetal cells from peripheral maternal blood using antibodies with high specificity and avidity. Methods: To generate antibodies, we purified nucleated red blood cells (NRBCs) from fetal livers and used them as the immunogen to generate monoclonal antibodies (mAbs) directed against surface antigens. Results: The four antibodies recognized at least two conformationally sensitive epitopes of the transferrin receptor. Isolation of NRBCs from 252 maternal blood samples using these antibodies in magnetic activated cell sorting after an initial density gradient centrifugation yielded 0-419 NRBCs per 25 mL of maternal blood. One antibody, 2B7.4, not only isolated the highest number of NRBCs (>10 in 90% of the samples) but also isolated these NRBCs in 78 consecutive maternal samples. Conclusion: Antibody 2B7.4 shows promise for the isolation of NRBCs from maternal blood and should allow studies concerning the source of these cells, fetal vs maternal, and the factors controlling their prevalence.

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