Figure 1.
Overview of the experimental approach. (A) Mouse is immunized at footpad with ricin A chain, and peripheral lymphoid organs, including bone marrow, spleen, and draining lymph node are isolated after 3 booster immunizations. (B) Total cells in bone marrow and spleen are collected, and VH and VL mRNA are reverse transcribed and amplified, which are used to construct scFv libraries, respectively. These are selected against ricin A chain using yeast surface display to isolate high-affinity binders. VH and VL cDNAs are also sequenced on an Illumina platform. (C) CD138+ antibody secreting cells are isolated from draining lymph node, and processed through our high-throughput VH:VL pairing platform.