Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Nov 29:17:14-20.
doi: 10.1016/j.csbj.2018.11.006. eCollection 2019.

Identity, Structure and Compositional Analysis of Aluminum Phosphate Adsorbed Pediatric Quadrivalent and Pentavalent Vaccines

Kristen Kalbfleisch  1 , Sasmit Deshmukh  1   2 , Carmen Mei  1 , Moriam Ore  1   3 , Wayne Williams  1 , Ibrahim Durowoju  1 , Jessica Duprez  1 , Sylvie Morin  3 , Bruce Carpick  1 , Marina Kirkitadze  1
Affiliations

Identity, Structure and Compositional Analysis of Aluminum Phosphate Adsorbed Pediatric Quadrivalent and Pentavalent Vaccines

Kristen Kalbfleisch et al. Comput Struct Biotechnol J. .

Abstract

Purpose: The goal of this study is to set an empirical baseline to map the structure-function relation of the antigens from the commercialized vaccine products.

Methods: To study the structural changes of protein antigens after adsorption several analytical tools including DLS, FTIR, Fluorescence, LD, and SEM have been used.

Results: All antigens have shown wide range of hydrodynamic diameter from 7 nm to 182 nm. Upon adjuvantation, the size distribution has become narrow, ranging from 10 to 12 μm, and has been driven by the derived diameter of aluminum phosphate (AlPO4) adjuvant. Further to examine size and morphology of adsorbed antigens, SEM has been used. The SEM results have demonstrated that the AlPO4 adjuvant suspension and adsorbed proteins consist of submicron particles that form a continuous porous surface. Diphtheria Toxoid (DT), Tetanus Toxoid (TT), and chemically-modified Filamentous Haemagglutinin (FHA) have shown surface adsorption to AlPO4. Secondary structure alpha-helix and beta-sheet content of DT and TT has increased after adsorption to AlPO4 adjuvant as shown by FTIR, whereas no significant changes were noted for other protein antigens. The results from Intrinsic Fluorescence have shown a structural rearrangement in DT and TT, consistent with the FTIR results. Multivalent vaccine product identity has been determined by FTIR as unique fingerprint spectrum.

Conclusion: The globular proteins such as DT and TT have shown changes in secondary structure upon adsorption to AlPO4, whereas fibrillar protein FHA has not been affected by adsorption. FTIR can be used as a lean technique to confirm product identity at different manufacturing sites.

Keywords: Adsorbed vaccines; FTIR; Fluorescence; Identity; Particle sizing; Protein conformation; SEM.

PubMed Disclaimer

Figures

Unlabelled Image
Graphical abstract
Fig. 1
Fig. 1
Particle size distribution of (a) pre-adsorbed and (b) adsorbed protein antigens by DLS and LD respectively. Representative traces include PRN (cyan), DT (red), FHA (blue), FIM (pink), TT (brown), and PT (grey) antigens. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 2
Fig. 2
Low vacuum SEM images of AlPO4 adsorbed antigens.
Fig. 3
Fig. 3
Representative FTIR spectra (top panels) and their calculated second derivatives (bottom panels) for all adsorbed (left panels) and pre-adsorbed (right panels) antigens. Some of measured spectra and calculated first derivative spectra are rescaled due to high signal intensity. Representative traces include PRN (cyan), DT (red), FHA (blue), FIM (pink), TT (brown), and PT (grey) antigens. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 4
Fig. 4
Overlay of FTIR spectra for Pediacel® (red trace), Pentacel® (blue trace) and Quadracel™ (black trace). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 5
Fig. 5
Intrinsic fluorescence emission spectra of DT (red), TT (brown), FHA (blue), and PRN (cyan) antigens in pre-adsorbed (solid traces) and adsorbed (short dashed traces) forms. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
See this image and copyright information in PMC

References

    1. Tung J.-S., Gimenez J., Przysiecki C.T., Mark G. Characterization of Recombinant Hepatitis B Surface Antigen using Surface Plasmon Resonance. J Pharm Sci. 1998;87(1):76–80. - PubMed
    1. Durowoju I., Bhandal K., Hu J., Carpick B., Kirkitadze M. Assessment of tertiary structure of protein vaccine antigens by differential scanning calorimetry. J Vis Exp. 2017;(121) doi: 10.3791/55262. http://www.jove.com/video/55262 - DOI - PMC - PubMed
    1. Vessely C., Estey T., Randolph T.W., Henderson I., Cooper J., Nayar R. Stability of a trivalent recombinant protein vaccine formulation against botulinum neurotoxin during storage in aqueous solution. J Pharm Sci. 2009;98:2970–2993. - PMC - PubMed
    1. Clapp T., Siebert P., Chen D., Jones Braun L.-T. Vaccines with aluminum-containing adjuvants: optimizing vaccine efficacy and thermal stability. J Pharm Sci. 2011;100(2):388–401. - PMC - PubMed
    1. Greiner V.J., Ronzon F., Larquet E., Desbat B., Estèves C., Bonvin J. The structure of HBsAg particles is not modified upon their adsorption on aluminium hydroxide gel. Vaccine. 2012;30(35):5240–5245. - PubMed