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Volume 112
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In-situ adsorption of aluminum-adjuvanted hepatitis B vaccine for optimized antigen distribution and enhanced vaccination
Zequn Liu a b c d, Zhe Lv c, Fan Yang c, Yufei Xia a b d *
a State Key Laboratory of Biopharmaceutical Preparation and Delivery, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, China
b University of Chinese Academy of Sciences, Beijing, 100049, China
c Sinovac Life Sciences CO., LTD., Beijing, 100085, China
d IPE-Sinovac Joint Laboratory of Advanced Adjuvant and Delivery Systems, Beijing, 100190, China
10.1016/j.partic.2026.03.001
Volume 112, May 2026, Pages 137-144
Received 4 January 2026, Revised 24 February 2026, Accepted 1 March 2026, Available online 13 March 2026, Version of Record 19 March 2026.
E-mail: yfxia@ipe.ac.cn
Highlights

• Antigen distribution varied with process and dictated vaccine immunogenicity.

• Multilayer adsorption was achieved by slow-titration in-situ adsorption process.

• In-situ adsorption process yielded high capacity alongside low strength.

• Optimal antigen distribution enhanced both stability and in vivo/vitro potency.


Abstract

Aluminum hydroxide (AH) exerts adjuvanticity predominately through efficient antigen loading. While extensive studies have reported how antigen adsorption onto pre-formed AH influences vaccine immunogenicity, the impact of in-situ antigen adsorption during AH formation remains underexplored. In this study, hepatitis B vaccines were prepared using conventional direct adsorption and two in-situ methods (rapid in-situ process and slow-titration in-situ formation), and compared in terms of physicochemical properties, antigen adsorption capacity and binding strength, antigen distribution, sedimentation properties and in vitro/in vivo potency. We showed that in-situ adsorption enabled multilayer antigen incorporation within the AH matrix, increasing adsorption capacity while reducing adsorption strength. Furthermore, slow-growth in-situ formation of AH promoted more extensive antigen layering than rapid-growth conditions, leading to sustained antigen release and superior in vivo efficacy. Together, these findings establish in-situ adsorption as an effective strategy for engineering antigen-AH interactions, enabling enhanced aluminum-adjuvanted vaccines without the need for additional adjuvant components.

Graphical abstract
Keywords
Hepatitis B vaccine; Aluminum adjuvant; In-situ adsorption; Antigen depot
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