Vector Labs Ab05151-23.0 Anti-Factor H Binding Protein [1A3], Rabbit IgG, kappa (100 μg) Centrifuge Bundle Savings at Sycamore Life Sciences

Description

Product Class: Purified
Clone ID: 1A3
Synonyms: fHbp; Genome-derived Neisseria antigen 1870; GNA1870; Lipoprotein 2086; LP2086
Amount: 100 μg
Application Codes Clone: ELISA; SPR; FC; Crystallography; Functional assays
Shipping Temperature: Wet Ice
Origin Pub PMID: 26106082
Buffer Composition: PBS with 0.02% Proclin 300.
Original Format: Fab
Storage Temperature: Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at -20⁰C.
Chimeric Use Statement: This chimeric rabbit antibody was made using the variable domain sequences of the original Human Fab format, for improved compatibility with existing reagents, assays and techniques.
Specificity Statement: This antibody is specific for the FHbp of Neisseria meningitidis serogroup B (variant group 1, ID 1) and exhibited no cross-reactivity with other FHbp variants tested—ID 4, 13, 74, 55 (variant group 1), ID 22, 77 (variant group 2), or ID 28 (variant group 3) (Beernink et al., 2015, PMID: 26106082; Sands et al., 2021, PMID: 34125873).
Application Notes: The original antibody format (recombinant human Fab, derived from subject A) was generated, and its specificity was verified using ELISA, demonstrating concentration-dependent binding to Neisseria meningitidis factor H binding protein (FHbp) ID 1 (variant group 1). It was used in inhibition ELISA with soluble FHbp variants, showing specific binding to FHbp ID 1 with no cross-reactivity to variants ID 4, 13, 74, 55 (variant group 1), ID 22, 77 (variant group 2), or ID 28 (variant group 3). Surface plasmon resonance (SPR) analysis revealed a binding affinity of K_D ≈ 0.44 nM to FHbp ID 1. Flow cytometry confirmed binding to live meningococci (strain H44/76, FHbp ID 1), with stronger binding at 10 µg/ml compared to 2 µg/ml. Flow cytometry also showed a slight enhancement (~2-fold) of human factor H (FH) binding to live meningococci in the presence of the Fab (10 µg/ml) (Beernink et al., 2015; PMID: 26106082). The antibody was expressed in mammalian cells and tested by ELISA, confirming concentration-dependent binding to FHbp variant group 1. Inhibition ELISA showed cross-reactivity with multiple FHbp variant group 1 sequences (91–100% sequence identity) but no reactivity with variant group 2 or variant group 3. ELISA also demonstrated ~2.8-fold enhancement of FH binding to purified FHbp at 5 µg/ml Fab. SPR analysis showed that the Fab reduced the association rate of FH to FHbp variant group 1, increasing the FH dissociation constant ~10-fold. X-ray crystallography (1.7 Å resolution, PDB: 7LCV) revealed binding to an epitope on the FHbp amino-terminal domain (buried surface area 916 Å²), adjacent to the FH binding site. Structural superposition modeling with an FH–FHbp complex suggested minimal direct Fab–FH contact but loop rearrangements in FHbp (Sands et al., 2021; PMID: 34125873).

Product Class:	Purified
Clone ID:	1A3
Synonyms:	fHbp; Genome-derived Neisseria antigen 1870; GNA1870; Lipoprotein 2086; LP2086
Amount:	100 μg
Application Codes Clone:	ELISA; SPR; FC; Crystallography; Functional assays
Shipping Temperature:	Wet Ice
Origin Pub PMID:	26106082
Buffer Composition:	PBS with 0.02% Proclin 300.
Original Format:	Fab
Storage Temperature:	Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at -20⁰C.
Chimeric Use Statement:	This chimeric rabbit antibody was made using the variable domain sequences of the original Human Fab format, for improved compatibility with existing reagents, assays and techniques.
Specificity Statement:	This antibody is specific for the FHbp of Neisseria meningitidis serogroup B (variant group 1, ID 1) and exhibited no cross-reactivity with other FHbp variants tested—ID 4, 13, 74, 55 (variant group 1), ID 22, 77 (variant group 2), or ID 28 (variant group 3) (Beernink et al., 2015, PMID: 26106082; Sands et al., 2021, PMID: 34125873).
Application Notes:	The original antibody format (recombinant human Fab, derived from subject A) was generated, and its specificity was verified using ELISA, demonstrating concentration-dependent binding to Neisseria meningitidis factor H binding protein (FHbp) ID 1 (variant group 1). It was used in inhibition ELISA with soluble FHbp variants, showing specific binding to FHbp ID 1 with no cross-reactivity to variants ID 4, 13, 74, 55 (variant group 1), ID 22, 77 (variant group 2), or ID 28 (variant group 3). Surface plasmon resonance (SPR) analysis revealed a binding affinity of K_D ≈ 0.44 nM to FHbp ID 1. Flow cytometry confirmed binding to live meningococci (strain H44/76, FHbp ID 1), with stronger binding at 10 µg/ml compared to 2 µg/ml. Flow cytometry also showed a slight enhancement (~2-fold) of human factor H (FH) binding to live meningococci in the presence of the Fab (10 µg/ml) (Beernink et al., 2015; PMID: 26106082). The antibody was expressed in mammalian cells and tested by ELISA, confirming concentration-dependent binding to FHbp variant group 1. Inhibition ELISA showed cross-reactivity with multiple FHbp variant group 1 sequences (91–100% sequence identity) but no reactivity with variant group 2 or variant group 3. ELISA also demonstrated ~2.8-fold enhancement of FH binding to purified FHbp at 5 µg/ml Fab. SPR analysis showed that the Fab reduced the association rate of FH to FHbp variant group 1, increasing the FH dissociation constant ~10-fold. X-ray crystallography (1.7 Å resolution, PDB: 7LCV) revealed binding to an epitope on the FHbp amino-terminal domain (buried surface area 916 Å²), adjacent to the FH binding site. Structural superposition modeling with an FH–FHbp complex suggested minimal direct Fab–FH contact but loop rearrangements in FHbp (Sands et al., 2021; PMID: 34125873).

Specifications

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Brand	Vector Labs
UNSPSC	12161500

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