Negative selection of chronic lymphocytic leukaemia cells using a bifunctional rosette-based antibody cocktail

BMC Biotechnology

Table 1 Comparison of the discussed four different enrichment approaches

	DGC	RS+DGC	DGC+MCS	DGC+FACS
Purity	ca. 70% ^a	ca. 93% ^a	> 98% ^d	90 – 95% ^f
	CD5⁺/CD19⁺	CD5⁺/CD19⁺	> 93% ^e	CD5⁺/CD19⁺
			CD19⁺
	86% (± 14%) ^b
	CD19⁺
	90% (± 7%) ^c
	CD5⁺/CD19⁺/CD23⁺
Time of procedure	< 60 mins.	< 90 mins.	> 120 mins (increasing with sample volume)	> 120 mins (increasing with sample volume)
Selection method	negative	negative	positive	positive
Limitations	highly depending on WBC count	dependent on WBC count when < 20 × 10⁶ cells/ml PB	long process time and expensive	long process time and very expensive
cost	low	medium	medium-high	high
Required Infra-structure	centrifuge	centrifuge	centrifuge + MCS Sorter	centrifuge + FACS sorter

DGC: density gradient centrifugation, RS+DGC: RosetteSep incubation prior to DGC, DGC+MCS: DGC and subsequent magnetic cell sorting, DGC+FACS: DGC and subsequent fluorescent activated cell sorting.
^a average purity experienced in our study
^b purity reported by Haslinger et al., 2004
^c purity reported by Falt et al., 2005
^d purity reported by Rosenwald et al., 2004 and Wiestner et al., 2003
^e Huttman et al., 2006
^f purity reported by Stankovic et al., 2004

ISSN: 1472-6750