Table 3. Hu225 pyrosequencing mutant analysis.
FACS Gate | CDR | total reads | WT | Mutant | indel (%) | 1 mutant (%) | > 1 mutant (%) |
---|---|---|---|---|---|---|---|
binding | H1 | 169,515 | 151,867 | 17,648 | 2,466 (14.0) | 14,627 (82.9) | 555 (3.1) |
“ | H2 | 157,383 | 51,265 | 106,118 | 14,361 (13.5) | 87,747 (82.7) | 4,010 (3.8) |
“ | H3 | 156,949 | 145,565 | 11,384 | 4,489 (39.4) | 6,667 (58.6) | 228 (2.0) |
“ | L1 | 185,058 | 72,533 | 112,525 | 15,704 (14.0) | 90,092 (80.1) | 6,729 (6.0) |
“ | L2 | 186,919 | 170,671 | 16,248 | 4,707 (29.0) | 11,338 (69.8) | 203 (1.2) |
“ | L3 | 183,699 | 84,961 | 98,738 | 50,269 (50.9) | 46,682 (47.3) | 1,787 (1.8) |
expression | H1 | 191,396 | 171,585 | 19,811 | 2,643 (13.3) | 16,665 (84.1) | 503 (2.5) |
“ | H2 | 180,492 | 91,927 | 88,565 | 14,153 (16.0) | 69,995 (79.0) | 4,417 (5.0) |
“ | H3 | 172,423 | 112,982 | 59,441 | 6,539 (11.0) | 49,653 (83.5) | 3,249 (5.5) |
“ | L1 | 205,999 | 121,315 | 84,684 | 12,715 (15.0) | 68,185 (80.5) | 3,784 (4.5) |
“ | L2 | 204,450 | 150,691 | 53,759 | 5,517 (10.3) | 46,805 (87.1) | 1,437 (2.7) |
“ | L3 | 204,621 | 103,969 | 100,652 | 44,681 (44.4) | 53,600 (53.3) | 2,371 (2.4) |
Totals | 2,198,904 | 1,429,331 | 769,573 | 178,244 (23.2) | 562,056 (73.0) | 29,273 (3.8) |
For each CDR identified the total number of reads is indicated along with the distribution of the reads between wild type and mutated and the further breakdown of mutant CDR sequences into frameshifts (indel), single mutant codons or more than one mutant codon.