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Fig. 3 | Inflammation and Regeneration

Fig. 3

From: A lesion-selective albumin-CTLA4Ig as a safe and effective treatment for collagen-induced arthritis

Fig. 3

Binding activities of mAlb-CTLA4Ig before and after MMP2/9 digestion. A mAlb-CTLA4Ig was digested with the indicated amount of MMP2/9 and then analyzed by western blot using an anti-mouse Fcγ antibody. Note that the major bands after digestion correspond to the size of conventional mCTLA4Ig (~ 55 kDa under reducing conditions). B Schematic representation of the cell-based ELISA used in the current study. C mAlb-CTLA4Ig were subjected to varying degrees of digestion by MMP2/9. Part of the digestion was analyzed by western blot to determine the degree of cleavage. The percent (%) cleaved Alb-CTLA4Ig was quantitated by the ImageJ software and is indicated below each lane. The other part of the digestion was added to the cell-based ELISA. The binding of CD80 to 1 nM conventional mCTLA4Ig was used as a positive control. D mAlb-CTLA4Ig was digested by MMP2/9. Graded amounts (10-fold serial dilution) of nondigested (blue curves) or MMP-digested (orange curves) mAlb-CTLA4Ig were tested for their binding kinetics in the cell-based ELISA. The binding kinetics of conventional mCTLA4Ig are shown as a black curve

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