Figure 5.

3H3 variable domain sequences described in this work. 3H3-Lv1/Hv1 represent the initial assembly of proteomic data into light and heavy V-domain sequences, where X represents ambiguous regions of sequence (gray highlighting = 113 Da; yellow = 158 Da; blue = 216 Da; green = 114 Da; magenta = 144 Da). An alignment of this draft to closest V and J translated germline gene segments is show below (a germline D-segment could not be assigned based on the initial draft sequence). 3H3-Lv2/Hv2 were refined draft sequences, as described in Table 1, and ambiguous regions in this light/heavy pair were programmed into 2 Fab phage display libraries. Further refinement after phage display led to 3H3-Lv2/Hv3, and finally 2 unique light/heavy pairings (3H3-Lv2/Hv4 and 3H3-Lv2/Hv5) that were part of recombinant Fabs. Sequence numbering is according to Kabat et al.¹⁸ CDRs are shown in bold and were defined according to the sequence definition of Kabat et al.,¹⁸ except for CDR-H1, which is the combined sequence and structural definition.¹⁹