Additionally, multiple stringent negative enrichments using targets to which the antibody fragment should not bind were selected against.
The clusters are named after significant enrichments of gene categories as determined by functional category enrichment using DAVID [41].
RBM44 was identified by proteomic analysis after intercellular bridge enrichment using TEX14 as a marker protein.
Blood was collected using the Vacutainer CPT Cell Preparation Tube System (BD, Heidelberg, Germany) and the blood samples were enriched in monocytes by negative selection, using RosetteSep Monocyte Enrichment Cocktail (StemCell Technologies, Vancouver, Canada).
Briefly, CD4+ T cells were isolated from spleens of 10- to 12-week-old, naïve, BALB/c mice by negative selection using CD4-enrichment columns (R&D System, Minneapolis, MN, USA).
For FACS analysis in Fig. 1, splenocytes were isolated and NK cells were enriched by negative selection using the SpinSep mouse NK cell enrichment kit according to the manufacturer's protocol (Stemcell Technologies, Meylan, France).
T cells were purified from collagenase-treated spleens by negative selection using T cell enrichment columns (R&D).
Either, CD4+ T cells or CD14+ cells were isolated by negative selection using the RosetteSep™ enrichment cocktail (StemCell Technologies, Vancouver, Canada).
The monocytes were isolated from PBMC by negative selection using the RosetteSep enrichment cocktail (Stemcell Technologies, Vancouver, BC, Canada).
Peripheral blood mononuclear cells (PBMCs) isolated from 10 RA patients (females, aged from 35 to 56 years) by density sedimentation on Ficoll-Hypaque gradients were separated immunomagnetically into T cells by negative selection using the RosetteSep enrichment cocktail according to the manufacturer's instructions (Stem Cell Technologies, Vancouver, BC, Canada).
