title: Novel serial positive enrichment technology enables clinical multiparameter cell sorting.
abstract: A general obstacle for clinical cell preparations is limited purity, which causes variability in the quality and potency of cell products and might be responsible for negative side effects due to unwanted contaminants. Highly pure populations can be obtained best using positive selection techniques. However, in many cases target cell populations need to be segregated from other cells by combinations of multiple markers, which is still difficult to achieve--especially for clinical cell products. Therefore, we have generated low-affinity antibody-derived Fab-fragments, which stain like parental antibodies when multimerized via Strep-tag and Strep-Tactin, but can subsequently be removed entirely from the target cell population. Such reagents can be generated for virtually any antigen and can be used for sequential positive enrichment steps via paramagnetic beads. First protocols for multiparameter enrichment of two clinically relevant cell populations, CD4(high)/CD25(high)/CD45RA(high) 'regulatory T cells' and CD8(high)/CD62L(high)/CD45RA(neg) 'central memory T cells', have been established to determine quality and efficacy parameters of this novel technology, which should have broad applicability for clinical cell sorting as well as basic research.
authors.affiliation: Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, Munich, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Stemberger
authors.forename: Stefan
authors.initials: S
authors.lastname: Dreher
authors.forename: Claudia
authors.initials: C
authors.lastname: Tschulik
authors.forename: Christine
authors.initials: C
authors.lastname: Piossek
authors.forename: Jeannette
authors.initials: J
authors.lastname: Bet
authors.forename: Tori N
authors.initials: TN
authors.lastname: Yamamoto
authors.forename: Matthias
authors.initials: M
authors.lastname: Schiemann
authors.forename: Michael
authors.initials: M
authors.lastname: Neuenhahn
authors.forename: Klaus
authors.initials: K
authors.lastname: Martin
authors.forename: Martin
authors.initials: M
authors.lastname: Schlapschy
authors.forename: Arne
authors.initials: A
authors.lastname: Skerra
authors.forename: Thomas
authors.initials: T
authors.lastname: Schmidt
authors.forename: Matthias
authors.initials: M
authors.lastname: Edinger
authors.forename: Stanley R
authors.initials: SR
authors.lastname: Riddell
authors.forename: Lothar
authors.initials: L
authors.lastname: Germeroth
authors.forename: Dirk H
authors.initials: DH
authors.lastname: Busch
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000032:analysis* / Q000276:immunology; D015704:CD4 Antigens / Q000032:analysis / Q000276:immunology; D016827:CD8 Antigens / Q000032:analysis / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000276:immunology; D053645:Interleukin-2 Receptor alpha Subunit / Q000032:analysis / Q000276:immunology; D019041:L-Selectin / Q000032:analysis / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000032:analysis / Q000276:immunology; D008297:Male; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology
chemical_list: D015703:Antigens, CD; D015704:CD4 Antigens; D016827:CD8 Antigens; D007140:Immunoglobulin Fab Fragments; D053645:Interleukin-2 Receptor alpha Subunit; D019041:L-Selectin; D017493:Leukocyte Common Antigens