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.