Three allergen LFIAs were developed for detecting peanut (PA) and
hazelnut (HA1 + HA2). Each assay has a different sandwich pair of
mAbs for their capture (T ) and detector carbon nanoparticle-labeled-antibody
(CNP-mAb), selected for their differences in sensitivity as observed
in prior works.24 (link)−26 (link) All assays used goat anti-mouse (GAMaB) IgG in PBS
(pH 7.6; 1.2 mg/mL; AffiniPure F(ab’)2 Fragment)
at theC line (Jackson Immunoresearch Laboratories Inc.,
Sanbio, Uden; The Netherlands) and were developed on nitrocellulose
membranes (140 CN; Unisart, Sartorius, Gottingen, Germany) overlaid
with an absorbent pad (Whatman, GE Healthcare, Eindhoven, The Netherlands)
and secured with a plastic backing (G and L, San Jose, CA, USA); seeSupporting Information Protocol S1 and S2 for
full details on CNP-mAb labeling and LFIA preparation details. For
the surface plasmon resonance (SPR) biosensor assay, an amine coupling
kit, pH scouting kit, HBS-EP buffer, and CM5 sensor chips were purchased
from GE Healthcare (Uppsala, Sweden); seeSupporting Information Protocol S3 for further details on the immobilization
procedure.
hazelnut (HA1 + HA2). Each assay has a different sandwich pair of
mAbs for their capture (
(CNP-mAb), selected for their differences in sensitivity as observed
in prior works.24 (link)−26 (link) All assays used goat anti-mouse (GAMaB) IgG in PBS
(pH 7.6; 1.2 mg/mL; AffiniPure F(ab’)2 Fragment)
at the
Sanbio, Uden; The Netherlands) and were developed on nitrocellulose
membranes (140 CN; Unisart, Sartorius, Gottingen, Germany) overlaid
with an absorbent pad (Whatman, GE Healthcare, Eindhoven, The Netherlands)
and secured with a plastic backing (G and L, San Jose, CA, USA); see
full details on CNP-mAb labeling and LFIA preparation details. For
the surface plasmon resonance (SPR) biosensor assay, an amine coupling
kit, pH scouting kit, HBS-EP buffer, and CM5 sensor chips were purchased
from GE Healthcare (Uppsala, Sweden); see
procedure.
