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    BIOTIN-ANALOG CONJUGATED ANTIBODIES FOR POSITIVE CELL SELECTION AND RELEASE
    3.
    发明专利
    BIOTIN-ANALOG CONJUGATED ANTIBODIES FOR POSITIVE CELL SELECTION AND RELEASE 未知

    公开(公告)号：CA2168205A1

    公开(公告)日：1995-03-16

    申请号：CA2168205

    申请日：1994-09-08

    Applicant: BAXTER INT

    Inventor： AL-ABDALY FAHAD A ,  CHANG YU-AN ,  GUILLERMO ROY ,  SCOUTEN WILLIAM H ,  SMITH ALAN K

    IPC: G01N33/53 ,  C07K16/28 ,  C07K16/30 ,  C12N11/00 ,  C12N15/02 ,  C12P21/08 ,  G01N33/543 ,  G01N33/569 ,  G01N33/58 ,  C12N5/00 ,  C07K1/13 ,  C12N5/08

    Abstract: The invention provides a stable conjugate of a target cell binding protein, preferably an antibody, covalently linked to a biotin analog moiety for use in positive cell selection and release. The invention also provides a method for forming a stable conjugate of an antibody and a biotin analog moiety comprising bringing an N-hydroxysuccinimide (NHS) ester of a biotin analog into reactive contact with an antibody in a medium with a pH of about 8 and at a temperature of about 2.degree.-8.degree.C. The invention also provides methods for the positive selection of target cells from a heterogeneous cell suspension. One method involves the formation within the cell suspension of a complex formed by the target cell bound to a primary antibody linked to a biotin analog moiety, a secondary anti-biotin antibody bound to the biotin analog moiety and linked to a cell separation means. The complex is separated from the heterogeneous cell suspension, then incubated with a competitive compound, preferably authentic biotin, for which the secondary antibody has a higher affinity than it has for the biotin analog. The competitive compound displaces the biotin analog moiety from the binding site of the secondary antibody, thus releasing the target cells from the complex. The cell separation means may then be separated from the target cells. Another method involves the formation within the heterogeneous cell suspension of a complex formed by the target cell bound to a primary antibody linked to a biotin analog moiety and a form of avidin, which is bound to the biotin analog moiety. The complex is separated from the heterogeneous cell suspension and then incubated with a competitive compound, preferably authentic biotin, for which avidin has a higher affinity than it has for the biotin analog. The competitive compound displaces the biotin analog moiety from the avidin binding site, thus releasing the target cells from the complex. The avidin may then be separated from the target cells. The invention also provides compositions of matter which are intermediate complexes in the above methods. One intermediate complex comprises a cell separation means bound to an anti-biotin antibody, which in turn is bound to a biotin analog moiety covalently linked to a target cell binding protein. Another intermediate complex comprises avidin bound to a biotin analog moiety covalently linked to a target cell binding protein.

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