Vector Labs Ab05152-23.0 Anti-Factor H Binding Protein [7B10], Rabbit IgG, kappa (100 μg) Centrifuge Bundle Savings at Sycamore Life Sciences

Description

Product Class: Purified
Clone ID: 7B10
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, 4, 13, 74, 55) and exhibited no cross-reactivity with FHbp variants 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 B) 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 cross-reactivity with FHbp ID 1, 4, 13, 74, and 55 (variant group 1) but no cross-reactivity with ID 22, 77 (variant group 2), or ID 28 (variant group 3). Surface plasmon resonance (SPR) analysis revealed a binding affinity (K_D ≈ 22.9 nM) to FHbp ID 1. Flow cytometry confirmed binding to live meningococci (strain H44/76, FHbp ID 1), with detectable binding at 10 µg/ml. Flow cytometry also showed 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). This 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 (87–100% sequence identity) but no reactivity with variant group 2 or variant group 3. ELISA also demonstrated ~4.5–10-fold enhancement of FH binding to purified FHbp at 5 µg/ml Fab. SPR analysis showed the Fab reduced both association and dissociation rates of FH to FHbp variant group 1, slightly increasing the FH dissociation constant. X-ray crystallography (2.0 Å resolution, PDB: 7KET; 1.5 Å for Fab alone, PDB: 7KE1) revealed binding to an epitope on the FHbp amino-terminal domain (buried surface area 817 Å²), adjacent to the FH binding site, with two salt bridges in the interface. 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:	7B10
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, 4, 13, 74, 55) and exhibited no cross-reactivity with FHbp variants 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 B) 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 cross-reactivity with FHbp ID 1, 4, 13, 74, and 55 (variant group 1) but no cross-reactivity with ID 22, 77 (variant group 2), or ID 28 (variant group 3). Surface plasmon resonance (SPR) analysis revealed a binding affinity (K_D ≈ 22.9 nM) to FHbp ID 1. Flow cytometry confirmed binding to live meningococci (strain H44/76, FHbp ID 1), with detectable binding at 10 µg/ml. Flow cytometry also showed 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). This 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 (87–100% sequence identity) but no reactivity with variant group 2 or variant group 3. ELISA also demonstrated ~4.5–10-fold enhancement of FH binding to purified FHbp at 5 µg/ml Fab. SPR analysis showed the Fab reduced both association and dissociation rates of FH to FHbp variant group 1, slightly increasing the FH dissociation constant. X-ray crystallography (2.0 Å resolution, PDB: 7KET; 1.5 Å for Fab alone, PDB: 7KE1) revealed binding to an epitope on the FHbp amino-terminal domain (buried surface area 817 Å²), adjacent to the FH binding site, with two salt bridges in the interface. 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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