Table 1.
Plasmid construction and yeast production level of VHH multimers.
| Yeast expression plasmida | Multimer VHH descriptiona | VHH name | Yeast VHH production levelb (mg/L) | Predicted MW (dalton) |
|---|---|---|---|---|
| pRL489 | SVT6-GS2-SVA12M2-H6 | T6A12 | 11 | 30,180 |
| pRL490 | SVT6-GS3-SVT16-L123Q-GS2-SVA12M2-H6 | T6T16A12 | 10 | 44,990 |
| pRL493 | SVT2-GS2-SVA12M2-H6 | T2A12 | 1 | 29,065 |
| pRL494 | SVT15-3FW4M-GS2-SVA12M2-H6 | T15A12 | 34 | 28,739 |
| pRL495 | SVT16-L123Q-GS2-SVA12M2-H6 | T16A12 | 99 | 29,192 |
| pRL496 | SVT2-GS2-SVG13M4-H6 | T2G13 | 17 | 27,968 |
| pRL497 | SVT6-GS2-SVG13M4-H6 | T6G13 | 1 | 29,083 |
| pRL498 | SVT15-3FW4M-GS2-SVG13M4-H6 | T15G13 | 41 | 27,643 |
| pRL499 | SVT16-L123Q-GS2-SVG13M4-H6 | T16G13 | 97 | 28,095 |
| pRL505 | SVT6-GS3-SVT15-3FW4M-GS2-SVA12M2-H6 | T6T15A12 | 3 | 44,537 |
| pRL506 | SVT15-3FW4M-GS3-SVT6-GS2-SVG13M5-H6 | T15T6G13 | 5 | 43,411 |
a
Nomenclature earlier used in patent application [45]. The name reflects the order of the protein domains present in the multimers from N- to C-terminus, where L123Q represents a mutation in (SV)T16 and 3FW4M represents 3 mutations in FW4 region of (SV)T15.
b
The production level in baker’s yeast was calculated from the yield of purified VHH from 0.5 L shaker flask cultures.