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. Author manuscript; available in PMC: 2018 Jan 16.
Published in final edited form as: Angew Chem Int Ed Engl. 2016 Dec 9;56(3):744–747. doi: 10.1002/anie.201608880

Figure 2. MPPD isolates high affinity and specificity aptamers.

Figure 2

(a) Specificity of aptamers derived from buffer-screen (left column) and serum-screen (right column), incubated with 1 nM labeled TNF-α either in buffer (top) or 10% human serum (bottom). Both pools exhibit high affinity without competitors (Quadrant IV, top row), but only the serum-screen pool retains high affinity in serum (bottom row). (b) Binding curves and calculated effective affinities to TNF-α for the top-performing serum-screen (S01) and bufferscreen (B01) aptamers as well as a previously published TNF-α aptamer (VR11), and a commercial TNF-α antibody (mAb11). In buffer, S01 shows slightly higher affinity than B01. In serum, S01 exhibits specific, high affinity exceeding that of mAb11, whereas B01 and VR11 show negligible binding. (c) S01 achieves a superior limit of detection to mAb11 in ELISA assays performed in serum. We also used MPPD to isolate (d) an NGAL aptamer with a KD of 0.92 nM in buffer and 1.28 nM in serum and (e) an HRP-2 aptamer with a KD of 0.13 nM in buffer and 0.12 nM in serum.