Lipid nanoparticles (LNPs) are widely used as delivery systems for mRNA vaccines. The stability and bilayer fluidity of LNPs are determined by the properties and contents of the various lipids used in the formulation system, and the delivery efficiency of LNPs largely depends on the lipid composition. For the quality control of such vaccines, here we developed and validated an HPLC-CAD method to identify and determine the contents of four lipids in an LNP-encapsulated COVID-19 mRNA vaccine to support lipid analysis for the development of new drugs and vaccines.

Background: As essential biomolecules composed of proteins and carbohydrate moieties, glycoproteins play pivotal roles in numerous biological processes. The glycosylation level plays a crucial role in determining the functionality of glycoproteins. Therefore, the precise quantification of glycan components in proteins holds significant importance for research on and development of polysaccharide–protein-conjugated vaccines. Methods: In this study, a novel glycan quantification approach was developed, leveraging analytical ultracentrifugation (AUC) technology that synergistically utilizes ultraviolet wavelength absorption and interference data to directly determine glycan mass fractions in glycoproteins. Results: This methodology expands the analytical framework for glycoproteins while retaining the intrinsic advantages of AUC, enabling analysis in native states with high reproducibility as indicated by low standard deviation across replicates. Conclusions: The approach was implemented in our proprietary AUC data analysis software called AUCAgent (v1.8.8), providing a new method for glycoprotein quantification and polysaccharide ratio determination in polysaccharide-protein-conjugate vaccines.

Antibody–drug conjugates (ADCs) are a pivotal technology for precision cancer therapy, harnessing the synergistic effects of antibody targeting and toxin delivery. However, traditional ADCs encounter limitations in efficacy that stem from tumor resistance, heterogeneity, and intense target competition. Dual-payload ADCs (DP-ADCs) represent a promising solution to these challenges, as they leverage dual mechanisms of action that mitigate acquired drug resistance and enhance adaptability to tumor heterogeneity. The complex structure of DP-ADCs presents substantial quality control hurdles. In this manuscript, we review the current payload selection and conjugation strategies of DP-ADCs and examine recent advances in quality control research. Specifically, we analyze the analytical challenges related to the quantification of free toxins, the determination of the total antibody content, and the characterization of the drug-to-antibody ratio and its distribution. Ultimately, the aim of this work is to provide valuable guidance for future DP-ADC quality control analyses to facilitate their clinical translation and application.

Antibody–drug conjugates (ADCs) are a pivotal technology for precision cancer therapy, harnessing the synergistic effects of antibody targeting and toxin delivery. However, traditional ADCs encounter limitations in efficacy that stem from tumor resistance, heterogeneity, and intense target competition. Dual-payload ADCs (DP-ADCs) represent a promising solution to these challenges, as they leverage dual mechanisms of action that mitigate acquired drug resistance and enhance adaptability to tumor heterogeneity. The complex structure of DP-ADCs presents substantial quality control hurdles. In this manuscript, we review the current payload selection and conjugation strategies of DP-ADCs and examine recent advances in quality control research. Specifically, we analyze the analytical challenges related to the quantification of free toxins, the determination of the total antibody content, and the characterization of the drug-to-antibody ratio and its distribution. Ultimately, the aim of this work is to provide valuable guidance for future DP-ADC quality control analyses to facilitate their clinical translation and application.

Background/Objectives: Bispecific antibody–drug conjugates (bispecific ADCs) are among the most rapidly advancing antibody therapeutic modalities, with only one product (BL-B01D1) currently approved for marketing worldwide to date. In this study, we investigated several critical quality attributes (CQAs) of a glyco-site-specific antibody–drug conjugate (ADC) targeting two distinct epitopes of HER2. Methods: We established quality control methods for the assessment of mispaired species, conjugation site, drug-to-antibody ratios (DARs), charge variants, residual free drugs, residual β1,4-galactosyltransferase I (GalT) enzyme, relative binding activity, and cell proliferation inhibition activity. Conclusions: An analytical method suite was developed for the bispecific ADC to comprehensively characterize and appropriately control its critical quality attributes, thereby guiding the research, development, and manufacturing process optimization of the ADC product and ensuring its safety, efficacy, and quality consistency.

Modern biotechnology has rapidly developed, and biotechnological drugs have become the center of global drug research and development as an important aspect of clinical treatment. These drugs have unique advantages, such as strong species specificity and prominent targeting, and they are widely used in the treatment of various intractable diseases. However, their complex molecular structure, poor stability, and significant heterogeneity make quality control much more difficult than that of traditional small-molecule drugs that require high-precision analytical methods. Analytical ultracentrifugation (AUC) technology was pioneered by Theodor Svedberg during the early 20th century, and it has become an indispensable biophysical tool through technological innovation. This technology has unique advantages for the quality control of biotechnological drugs, including non-destructive detection, high resolution, and wide applicability. Therefore, AUC has been extensively adopted for the characterization of various biological products. This review systematically summarizes AUC technology applications in major categories of biotechnological drugs, with the aim to provide technical guidance and promote the standardized application of AUC.