Anti-Factor H Binding Protein [1A3], Human Fab fragment, His-Tagged, kappa

Catalogue Number: AB05151-10.29-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):	100 ug
Host name:	Human
Clone:	1A3
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. 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 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 KD ~ 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).

AB05151-10.29 Datasheet

Price

£525.00

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