Anti-Factor H Binding Protein [7B10], Human Fab fragment, His-Tagged, kappa

Catalogue Number: AB05152-10.29-BT-ABA

Manufacturer:	Vector Laboratories, Inc (ABA)
Type:	Recombinant Monoclonal
Alias:	fHbp; Genome-derived Neisseria antigen 1870; GNA1870; Lipoprotein 2086; LP2086
Shipping Condition:	Blue Ice
Unit(s):	500 ug
Host name:	Human
Clone:	7B10
Isotype:
Immunogen:	The original antibody was generated by immunizing human volunteers with a multicomponent serogroup B meningococcal vaccine containing recombinant FHbp corresponding to a sequence from a serogroup B Neisseria meningitidis strain.
Application:	ELISA, FC, FA, SPR, Crstapy

Overview
Datasheets

Additional Text

Purification

Purified

Antibody Clonality

Recombinant Monoclonal

Storage Note

Store at 4⁰C for up to 3 months. Note, this antibody is provided without added preservatives, it is therefore recommed this antibody be handled under sterile conditions. For longer storage, aliquot and store at -20⁰C.

Short Description

This reformatted human antibody was made using the variable domain sequences of the original Human Fab format, for improved compatibility with existing reagents, assays and techniques.

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 (KD ~ 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).

AB05152-10.29-BT Datasheet

Price

£2,215.00

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