Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5968753

Title:Positive and positive/negative cell selection mediated by peptide release
Inventors:Tseng-Law, Janet; Whitter, CA, USA
Kobori, Joan A.; Pasadena, CA, USA
Al-Abdaly, Fahad A.; Torrance, CA, USA
Guillermo, Roy; Carson, CA, USA
Helgerson, Sam L.; Pasadena, CA, USA
Deans, Robert J.; Claremont, CA, USA
Summary:Described herein are short peptides and methods of their use in the displacement of antibodies from cell surface antigens. The invention relates to a non-enzymatic method for the release of cells which have been positively selected from a heterogeneous cell suspension by antibody-mediated binding to beads or to other solid supports, which results in the formation within the cell suspension of a complex comprising the solid support linked to a primary antibody, which in turn is specifically bound to a cell surface antigen on the target cells. Further disclosed are methods of positive/negative cell selection, methods for the identification of specific peptides for the release of target cells from the binding of primary antibodies, and methods for the purging of undesired tumor cells or lymphocytes from the positively selected cell population. Therapeutic applications include the use of purified CD34+ cell compositions for reinfusion to a cancer patient after high-dose chemotherapy in order to reconstitute the patient's immune system.
Abstract:The invention provides a non-enzymatic method for the release of cells which have been positively selected from a heterogeneous cell suspension by antibody-mediated binding to beads or other solid support. The method entails forming within the cell suspension a complex comprising the solid support linked to a primary antibody, which in turn is specifically bound to a cell surface antigen on the target cells. The complex is separated from the cell suspension, and then contacted with a specific peptide which binds to the primary antibody, displacing the antibody from the cell surface antigen, thereby releasing the target cell from the complex. The invention also provides methods for positive/negative cell selection wherein target cells having a first antigen are selected from a heterogeneous cell suspension containing undesired cells having a second antigen. The invention also provides methods for identifying a specific peptide useful for the release of a target cell from the binding of a specific primary antibody. The methods of the invention are particularly useful for the positive selection of CD34+ hematopoietic stem cells and the concomitant purging of undesired tumor cells or lymphocytes from the positively selected cell population. The purified CD34+ cell composition is then useful for reinfusion to a cancer patient after high-dose therapy in order to reconstitute the patient's immune system.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5968753
Application Number:US1995000482228
Date Filed:07/06/1995
Date Published:19/10/1999
Assignee:Nexell Therapeutics, Inc., Irvine, CA, USA


 
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