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Figure 3 | BMC Biotechnology

Figure 3

From: Incorporation of albumin fusion proteins into fibrin clots in vitro and in vivo: comparison of different fusion motifs recognized by factor XIIIa

Figure 3

FXIIIa-mediated cross-linking of recombinant proteins and plasma-derived α 2 AP to biotinylated pentylamine. Panel A shows transglutamination reactions, in which 20 nM FXIIIa cross-linked 5 mM BPA to 1.7 μM HSA fusion proteins (identified below the × axis), were stopped after 2.5 minutes and the relative BPA incorporation was quantified using anti-HSA capture and alkaline phosphatase-conjugated streptavidin detection as outlined in "Methods". Colour generation was quantified as the optical density at 405 nm (OD (405 nm)). The mean ± standard deviation (SD) of 6 determinations is shown, with background (reactions lacking FXIIIa) subtracted. Panel B shows a time course of FXIIIa-mediated cross-linking of BPA to 1.7 μM α2AP, using the same conditions as employed in Panel A except that anti-α2AP antibodies were used to capture biotinylated α2AP after reaction termination at various time points. The mean ± the standard deviation (SD) of 3 determinations is shown, with background (reactions lacking FXIIIa) subtracted. Panel C shows BPA-α2AP crosslinking reactions of 5 minutes' duration, as shown in panel B, except that competitor proteins identified on the × axis were included at 12 μM. The mean ± the SD of 5-14 determinations is shown, with background (reactions lacking FXIIIa) subtracted.

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BMC Biotechnology

ISSN: 1472-6750

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