SDS-PAGE, ELISA and biacore analysis of anti-cTnI mAb, 20B3 (Fig.1-3)
Fig. 1 SDS-PAGE and Western Blot analysis of purified anti-cTnI mAb.
The SDS-PAGE containing purified anti-cTnI mAb bands (light chain: ~20 kDa; heavy chain ~50 kDa). The Western blot shows the specific band on Western blotting (~23kDa) of the cTnI antigen probed with the anti-cTnI scFv. PR: Pre-stained Page Ruler; cTnI: cTnI Antigen. Figure 1b shows the schematic diagram of Western blotting analysis.
Fig. 2 Titrations of purified the 20B3 and commercial Hytest 228 anti-cTnI mAbs.
Reciprocal antibody dilutions in 1 % (w/v) PBSTM titrated against cTnI-coated onto the surface of a microtitre plate. The purified mAb titred in a similar fashion to the commercial standard and indicated no adverse loss of activity due to the purification strategy.
Fig. 3 Kinetic analysis of interactions between anti-cTnI mAb 20B3 and cTnI using the Biacore 3000 system.
The coloured lines represent the response data of various quantities of cTnI. The black lines represent the local fit to the response data shown as coloured lines.