Research Products

HBsAgGi: HB surface antigen glycan isomer

A monoclonal antibody to O-glycosylated PreS2 on M-HBs, which is predominantly expressed by DNA-containing viral particle.

Overview:

HBV envelope protein is composed of three types of surface antigens, S-, M-, and L-HBs, which are produced from an HBsAg gene containing PreS1, PreS2 and S-domains1. HBsAgs are heavily glycosylated with N-glycan and O-glycan2,3. Whole glycan structural analyses revealed that PreS2 domain on M-HBs, but not on L-HBs, contains highly conserved O-glycosylated site in genotype C (gC)4,5.

A recombinant monoclonal antibody against HBsAg glycan isomer (HBsAgGi) was generated by using O-glycosylated PreS2 peptide. Compared to traditional HBsAg-testing which recognizes all viral particles, HBsAgGi specifically recognizes infectious HBV particles (DNA virion).

HBsAgGi

Recombinant mouse IgGI, 50μg


US$700-

 Isotype: mouse IgG1
 Light chain: kappa
 Amount: 50 μg
 Storage:Store at 4°C for short term. Upon delivery store at  -20°C for long term and avoid freeze / thaw cycle.

Applications:
 1. Western
 2. Immunoprecipitation/Separation of HBV particles
 3. ELISA
 4. HBV infection inhibition assay
 5. Immunohistochemistry

HBsAgGi specifically recognizes M-HBsAg in HBV virion

HBV virion contains envelope protein (HBsAg), HBV core and HBV DNA. HBsAg gene consists of PreS1 (blue), PreS2 (red), and S (black) -domains. About half of S-HBsAgs is modulated by N-glycan and the rest is non-glycosylated. PreS2 domain in M-HBsAg is highly O-glycosylated, but not in L-HBsAg. S-HBsAg is the most abundant and forms non-infectious particles. Infectious HBV particles consist of all HBsAgs and contains HBV DNA. HBsAgGi recognizes O-glycosylated PreS2 domain in M-HBsAg of genotype C.

HBsAgGi specifically recognizes M-HBsAg in HBV virion

HBsAgGi antibody vs S antibody in recognition of HBV particles

In HBV patients’ serum, non-infectious particles are much dominant than infectious particles containing HBV DNA. Anti-S antibody may recognize all HBV particles including non-infectious particles, However, HBsAgGi specifically recognizes HBV particles containing M-HBsAg, resulting in efficient recognition of infectious HBV particles. HBsAgGi measuring will classify HBV patients differently from the S-antibody measuring.

HBsAgGi antibody vs S antibody in recognition of HBV particles

HBsAgGi and other markers to detect infectious HBV

Comparison of HBV markers. Measuring qHBsAg and HBV DNA have been used, while HBcrAg and HBV RNA are new markers to diagnose patient condition. Target HBV particles or molecules are indicated in the table. Circle (〇) indicates the target particle and molecules, and X indicates that the marker cannot recognize or not recommended.
HBsAgGi detects M-HBsAg-containing particles such as DNA or RNA virion.

HBsAgGi and other markers to detect infectious HBV

Examples:

HBsAgGi

Western blotting using HBsAgGi (Left) and PreS2 antibody (Right)

  • Left: HBsAgGi-gC antibody, Right: anti-PreS2 antibody
  • Lane 1: S-HBsAg 50 ng
  • Lane 2: M-HBsAg 50 ng
  • Lane 3: L-HBsAg 50 ng
  • Secondary antibody: HRP -labeled anti-mouse IgG
  • HBsAgGi-gC

    ELISA using HBsAgGi-gC coated plate and M-HBsAg as a standard sample

  • Standard material: M-HBsAg (3.7 – 900 ng/mL)
  • Tested samples: sera from patients with HBV (2 μL)
  • Serum #1: HBsAg 6610 IU/mL, HBV DNA 3.1
  • Serum #2: HBsAg 8612 IU/mL, HBV DNA 0
  • Serum #3: HBsAg 36600 IU/mL, HBV DNA 8.3
  • Detection: Biotinylated HBsAgGi-gC antibody and HRP-labeled streptavidin
  • HBsAgGi-gC

    Immunoprecipitation using HBsAgGi-gC

  • Original sample (Orig): HBV patient serum (4.1 logIU/20 μL)
  • IP condition: Biotinylated HBsAgGi-gC 2 μg, Streptavidin-conjugated magnetic beads 10 μL/1 mL TBS-T, 4°C, 16 hours
  • Quantification of HBV DNA: HBV DNA per 20 μL from precipitated fraction (IP) and supernatant fraction (Sup) was measured by real-time PCR method.
  • References:

    1. Schadler and Hildt (2009) Viruses 1:185-209.
    2. Schmitt et al. (2004) J Gen Virol 85:2045-2053.
    3. Dobrica et al. (2020) Cells 9:1404.
    4. Wagatsuma et al. (2018) Anal Chem 90:10196-10203.
    5. Angata et al. (2021) Biochim Biophys Acta Gen Subj.  1866:130020

    HBsAgGi

    Recombinant mouse IgGI, 50μg

    US$700-

    RCMG Inc.


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