Table 2. Sequences of the RGYW-mutant scFv obtained after parasite selection.
| ScFv ID | Nucleotide sequences | Amino acid mutations | |
|---|---|---|---|
| Parental Ab—TG130 | AGT Act | Ser Thr |  |
| Alanine mutant—RA15 | AGG Gct | Arg Ala |  |
| CGC Gct | |||
| CGG Gct | |||
| RT51 | AGG Act | Arg Thr |  |
| CGC Act | |||
| RS55 | AGG Tct | Arg Ser |  |
| CGG Tct | |||
| LT3 | CTG Act | Leu Thr |  |
| CTC Act | |||
Note:
Affinity maturation of scFv TG130 at RGYW hot spot motifs resulted in site-directed mutagenesis at amino acid positions 27 and 28 of VL CDR1. The unique sequences of the mutant scFv antibodies isolated from biopanning selection against T. gondii are listed, along with the parental antibody. The mutated amino acid residues are color-coded according to its side-chain properties—Uncharged Polar (Red), Hydrophobic (Blue), Charged Polar (Green).