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. 2014 Feb 5;10(4):1013–1023. doi: 10.4161/hv.27753

Table 2. In vitro relative potency assays of Hepatitis B vaccine reported in the literature and proposed assays based on the results in this report3.

Method Key reagent Information
on epitopea
Information on disulfide- dependent epitope Comment Country Reference
Sandwich ELISAb (Bead-based) mAbs + No Discontinued in 2005 USA Auszyme kit. Schofield et al. 200222
Inhibition ELISAc pAb - No One site assayd Brazil Cardoso et al. 200135
Cuba Cuervo et al. 200421
Cuba Cuervo et al. 200827
Inhibition ELISAe pAb and mAb + No One site assayd Belgium Giffroy et al. 200636
Sandwich ELISAf pAb as capture Ab mAb as detection Ab + No mAb affinity was determination India Shanmugham et al. 201037
Sandwich ELISAg mAbs as capture Ab pAb as detection Ab - No   Iran Karimzadeh et al. 201038
Sandwich ELISAh mAbs + No   Brazil Costa, et al. 201126
Sandwich ELISAi mAbs     Multifaceted, highly specific, and sensitive China This work
5F11 as detection Ab ++ Yes
42B6 as detection Ab + No

a The “+” represents that method aim at specific epitope on HBsAg, “-” otherwise. The “++” means against specific epitope, furthermore, monitoring the HBsAg conformational changes due to changes in disulfide bonds. bMethod was based on AuszymeTM kit, a bead based assay with anti-HBsAg and mAbs. (http://www.fda.gov/downloads/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/LicensedProductsBLAs/BloodDonorScreening/InfectiousDisease/ucm077519.pdf). cPlates were coated HBsAg polyclonal antibodies.The quantity of non-neutralized antibodies was then measured via HBsAg-HRP conjugated. dThe assays were potentially useful in monitoring the vaccine antigenicity on adjuvants. ePlated were coated HBsAg polyclonal antibodies, non-neutralized mAb was used compete Ab for the method, then the quantity of antibodies were measured via goat anti-human IgG antibodies labeled with peroxidase. fPlates were coated polyclonal guinea pig anti-HBsAg antibody and mouse anti-HBsAg antibody was for measuring HBsAg. gPlates coated 2 monoclonal antibodies and a polyclonal antibody conjugated with biotin was used for detect Ab. hTwo human monoclonal antibodies were used in this method, one was coated Ab and the other for detecting. iIn this paper, we proposed a pair of assays (sandwich conformational / sandwich linear) for monitoring our HBsAg. The assay has advantage of detecting conformational changes by an “antigenicity ELISA” with mAb which is highly sensitive to the disulfide bond status and quantifying HBsAg by a “mass ELISA” with mAb recognizing a linear epitope.