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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: Methods. 2013 Jun 26;64(2):10.1016/j.ymeth.2013.06.018. doi: 10.1016/j.ymeth.2013.06.018

Figure 1.

Figure 1

Outline of the methodology for the combined screening of the antigen specificity and phenotype of single auto-reactive B cells. B cells obtained by enrichment from CCP+ donor derived PBMCs are loaded onto a nanowell array. Microengraving is used to determine CCP reactivity and automated fluorescence microscopy is employed to determine the phenotype of the B cells, on-chip. Post data-analysis, the memory B cells secreting CCP-specific hmAb are retrieved by micromanipulation. Single cell RT-PCR is performed to enable the amplification of the Ig VH and VL regions.