Fig. 3: Biolayer interferometry antibody screen (BIAS) scheme and control experiments. | Communications Biology

Fig. 3: Biolayer interferometry antibody screen (BIAS) scheme and control experiments.

From: Rare antibody phage isolation and discrimination (RAPID) biopanning enables identification of high-affinity antibodies against challenging targets

Fig. 3

a BIAS scheme. Representative biolayer interferometry (BLI) sensograms of each distinct outcome (Hit, False positive, Negative) are shown. (i) Association-1: Biological tips bound with Ag are transferred to crude PPEs induced for expression, (ii) Association-2: tips are transferred to wells containing identical PPE sample from Association-1 + anti-myc IgG, (iii) Dissociation: tips from (ii) are transferred to wells with no PPE and no IgG. The Association-2 step distinguishes hits versus false positives. b–d BLI sensograms of BIAS performed with ARS1620 and P1A4 spiked samples. b P1A4 (250 nM) spiked in PBS. A signature linear curve is observed in Assco-2 where the anti-myc antibody (9E10) is present, while this is not observed with the control with no 9E10. c P1A4 (250 nM) spiked in TG1-PPE. Results are similar as (c). d Concentration series of P1A4 spiked in TG1-PPE (10–300 nM).

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