UNASSIGNED: Fish @entity566 is associated with moderate to severe IgE-mediated reactions to the @entity460 binding parvalbumins present in fish muscle. @entity566 to multiple fish species is caused by @entity15537 -specific cross-reactive IgE recognizing conserved epitopes. In this study, we aimed to produce cross-reactive single chain variable fragment ( @entity10390 ) antibodies for the detection of parvalbumins in fish extracts and the identification of IgE epitopes. @entity15537 -specific phage clones were isolated from the @entity1 ETH-2 phage display library by three rounds of biopanning either against cod @entity15537 or by sequential biopanning against cod (Gad m 1), @entity7136 (Cyp c 1) and @entity5660 (Onc m 1) parvalbumins. While biopanning against Gad m 1 resulted in the selection of clones specific exclusively for Gad m 1, the second approach resulted in the selection of clones cross-reacting with all three parvalbumins. Two clones, @entity10390 -gco9 recognizing all three parvalbumins, and @entity10390 -goo8 recognizing only Gad m 1 were expressed in the @entity526 non-suppressor strain HB2151 and purified from the periplasm. @entity10390 -gco9 showed highly selective binding to parvalbumins in processed fish products such as breaded cod sticks, fried @entity7136 and smoked trout in Western blots. In addition, the @entity10390 -gco9-AP produced as alkaline phosphatase fusion protein, allowed a single-step detection of the parvalbumins. In competitive ELISA, @entity10390 -gco9 was able to inhibit binding of IgE from fish @entity56 @entity1 ' sera to all three b-parvalbumins by up to 80%, whereas inhibition by @entity10390 -goo8 was up to 20%. 1H/15N HSQC NMR analysis of the rGad m 1: @entity10390 -gco9 complex showed participation of @entity3689 residues conserved among these three parvalbumins explaining their cross-reactivity on a molecular level. In this study, we have demonstrated an approach for the selection of cross-reactive @entity15537 -specific antibodies that can be used for allergen detection and for mapping of conserved epitopes.

A Cross-Reactive XXXX Single-Chain Antibody for Detection of Major Fish Allergens, Parvalbumins, and Identification of a Major IgE-Binding Epitope.

@entity1 :: ('9606', 'Species') :: ['human', 'patients'] @entity5660 :: ('8022', 'Species') :: ['rainbow trout'] @entity566 :: ('MESH:D006967', 'Disease') :: ['allergy', 'Allergy'] @entity56 :: ('MESH:D004342', 'Disease') :: ['allergic'] @entity460 :: ('MESH:D002118', 'Chemical') :: ['calcium'] @entity10390 :: ('652070', 'Gene') :: ['scFv'] @entity7136 :: ('7962', 'Species') :: ['carp'] @entity3689 :: ('D000596', 'Chemical') :: ['amino acid'] @entity15537 :: ('5816', 'Gene') :: ['parvalbumin', 'Parvalbumin'] @entity526 :: ('562', 'Species') :: ['E. coli']
@entity1 :: (9606,Species) :: ['Human']
