Fig. 1. DNA gel analysis of DR4, Streptavidin, hAY4 ScFv and hAY4 ScFv-STR genes cloned into pUC 119 expression vector.
Fig. 2. Plasmid construction of Streptavidin, DR4, hAY4 ScFv and hAY4 hAY4-STR.
Fig. 3. SDS-PAGE analysis of soluble proteins expressed in E. coli.
Fig. 4. SDS-PAGE analysis of Streptavidin and hAY4 ScFv-STR using heating or non-heating condition to determine non-covalent tetramerization effect.
Fig. 5. Size exclusion chromatography (SEC).
Fig. 6. Ouchterlony assay and ELISA analysis to determine Biotin binding activity of Streptavidin and hAY4 ScFv-STR.
Fig. 7. ELISA analysis and Westernblot to determine antigen binding activity of hAY4 ScFv-STR.
Fig. 8. Surface plasmon resonance (SPR) analysis of hAY4 ScFv, STR and hAY4 ScFv-STR.
Fig. 9. 3D modeling of hAY4 ScFv-STR tetramer.
Table 1. Primers used for PCR amplification of DR4, Streptavidin, hAY4 ScFv, and hAY4 ScFv-Streptavidin genes
Table 2. Kinetic binding parameters for the interactions of hAY4 ScFv and hAY4 ScFv-STR with DR4 determined by SPR analysis
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