Recombinant Mouse Anti-Neuraminidase Antibody Fab Fragment (NC10scFv(15))

ScFv-0 shows two lower molecular mass bands of ~14 and 15 kDa (arrowed) corresponding to the VH and VL domains produced by proteolytic cleavage of the scFvs during isolation as described in the text. The far right lane shows the monomer peak (Fv) isolated from the scFv-0 preparation (left lane) by gel filtration.

Kortt, A. A., Lah, M., Oddie, G. W., Gruen, C. L., Burns, J. E., Pearce, L. A., ... & Webster, R. G. (1997). Single-chain Fv fragments of anti-neuraminidase antibody NC10 containing five-and ten-residue linkers form dimers and with zero-residue linker a trimer. Protein engineering, 10(4), 423-433.

(a) Shows the scFv-15 containing monomer, dimer and some higher Mr multimers, (b) shows the scFv-10 with predominantly dimer and (c) shows the scFv-0 eluting as a single peak with an Mr of ~70 kDa. The column was equilibrated with PBS, pH 7.4, and run at a flow rate of 0.5 ml/min.

Kortt, A. A., Lah, M., Oddie, G. W., Gruen, C. L., Burns, J. E., Pearce, L. A., ... & Webster, R. G. (1997). Single-chain Fv fragments of anti-neuraminidase antibody NC10 containing five-and ten-residue linkers form dimers and with zero-residue linker a trimer. Protein engineering, 10(4), 423-433.

An injection volume of 30 ml and a flow rate of 5 ml/min were used. For the NC10 scFv-15 monomer the baseline response returned to the starting value after 6000 s and NC10 scFv-15 monomer at a series of concentrations (15–231 nM) was run to measure the rate constants for this interaction.

Kortt, A. A., Lah, M., Oddie, G. W., Gruen, C. L., Burns, J. E., Pearce, L. A., ... & Webster, R. G. (1997). Single-chain Fv fragments of anti-neuraminidase antibody NC10 containing five-and ten-residue linkers form dimers and with zero-residue linker a trimer. Protein engineering, 10(4), 423-433.