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Antibodies binding to CD34+/CD36+ fetal but not to adult cells

A fetal cell, cell antigen technology, applied in the field of antibodies that bind to fetal but not adult CD34+/CD36+ cells

Inactive Publication Date: 2007-07-18
THE BOARD OF TRUSTEES OF THE UNIV OF ILLINOIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A particular limitation is that the relevant maternal and fetal cells used for this purpose are primarily stem cells and cells in the BFU and CFU stages of differentiation

Method used

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  • Antibodies binding to CD34+/CD36+ fetal but not to adult cells
  • Antibodies binding to CD34+/CD36+ fetal but not to adult cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0044] Example 1: Preparation of fetal liver cells

[0045] Fetal liver cells can be obtained from Cambrex Bio Science, Walkersville, M.D. Alternatively, it can be isolated from fetal livers (FL, 15-22 weeks of gestation) from 5 different donors, as described below.

[0046] Fetal livers from five donors were homogenized in DPBS / 0.2% BSA solution and passed through a metal sieve. Among them, DPBS does not contain Ca ++ / Mg ++ Dulbecco's phosphate buffered saline solution (Biowhittaker, Walkersville, MD); BSA is bovine serum albumin (Sigma St. Louis, MO), and contains 50 μg / ml gentamicin sulfate (Life Technologies, Grand Island NY). CD34 + Cells can be sorted from the homogenate using MACS or FACS as described in Example 3. Or, in accordance with the MACS or FACS method described in Example 3 to sort CD34 + Before the cells, the homogenate was further purified as described below.

[0047] Use immunomagnetic microspheres to exclude mature red blood cells and other lineage-positive c...

example 2

[0048] Example 2: Preparation of mobilized peripheral blood cells

[0049] The mobilized peripheral blood cells are obtained from Cambrex, and can also be obtained from the following methods.

example 3

[0052] Example 3: Adult and fetal CD34 + Purification of progenitor cells

[0053] The fetal liver cells and MPB mononuclear cell fractions of Example 1 and Example 2 were recorded in CD34 according to any of the following methods. + Cell enrichment.

[0054] MACS method

[0055] Fetal liver cells and MPB monocytes were prepared according to the methods of Examples 1 and 2, respectively, and MACS CD34 was used + The separation kit (Miltenyi Biotec, Auburn, CA) is in accordance with the instructions for use in CD34 + Cellular immune magnetic beads are enriched. Briefly, monocytes are incubated with hapen-labeled anti-CD34 antibody (QBEND-10, BD Pharmingen, San Diego, CA) in 0.1% human IgG (Bayer Elkhart, IN) as a blocking agent, and then ligated with Co-culture to MACS microbeads against hapen. The labeled cells are filtered through a 30μm nylon mesh to remove cell clumps and aggregates. Then labeled CD34 + The cells are captured from the mixture through a high gradient magnetic se...

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Abstract

This invention relates to antibodies that specific bind to fetal CD36+ cells in preference to binding to maternal CD36+ cells and methods for using these antibodies to detect and separate fetal cells from adult biological fluids including maternal peripheral blood.

Description

[0001] This application claims the priority of U.S. Patent Application No. 60 / 579,693 filed on June 14, 2004 and U.S. Patent Application No. 60 / 618,963 filed on October 15, 2004. The contents of the above-mentioned patent applications are expressly referenced The method is incorporated into this article. Technical field [0002] The present invention relates to fetal CD36 + Cells instead of maternal CD36 + Cell-bound antibodies, and the use of these antibodies to detect and isolate fetal cells from maternal peripheral blood. Background technique [0003] The discovery of fetal red blood cells in maternal blood after childbirth was reported by Zipursky in 1959 ("Foetal erythrocytes in the maternal circulation," Lancet 1:45 1-52). Later, it was discovered that fetal red blood cells, lymphocytes and trophoblast cells were present in the maternal blood during pregnancy (Herzenberg et al., 1979, Proc Natl Acad Sci USA, 76: 1453-1455; Bianchi et al., 1990, Natl Acad Sci USA, 87: 3279-83...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12Q1/68G01N33/12
Inventor 阿伦·夏尔马舍曼·伊莱亚斯
Owner THE BOARD OF TRUSTEES OF THE UNIV OF ILLINOIS
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