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. 2016 Jun 15;6:27985. doi: 10.1038/srep27985

Figure 3. Expression, purification and antigen binding kinetics of chimeric or humanized antibody.

Figure 3

(a) SDS-PAGE and (b) Western Blot assay of stable expressed and purified HCLC/H3L2. Lane 1 and 3 were non-reduced and reduced HCLC, lane 2 and 4 were non-reduced and reduced H3L2 respectively. Lane M were marker with molecular weight of 250, 130, 100, 70, 55, 35, 25 KDa. (c,d) Antigen binding kinetics analyzed by SPR. Sensorgrams of series concentrations of rhDLL4 binding to (c) HCLC or (d) H3L2 were recorded and fitted with a 1:1 binding model. kd and ka of HCLC were 2.42 × 10−5 1/s and 1.72 × 107 1/Ms, and KD calculated from kd/ka was 1.14 × 10−12 M. R2 value of HcLc was 1.44 and Rmax was 50.77 RU. kd and ka of H3L2 were 2.59 × 10−5 1/s and 1.15 × 107 1/Ms, and KD was 2.26 × 10−12 M. R2 value of H3L2 was 1.68 and Rmax was 50.9.