biotinylated mal ii  (Vector Laboratories)

Journal: Frontiers in Pharmacology

Article Title: Mycobacterium tuberculosis infection drives osteoclast overactivation via α2,3-Sialylation to promote pathological bone destruction

doi: 10.3389/fphar.2026.1738896

Figure Lengend Snippet: α2,3-sialylation is required for BCG-induced osteoclast differentiation and activity (A) Comparison of differentially expressed genes (DEGs) between BCG-infected osteoclasts (RB) and uninfected controls (R). (n = 3 biological replicates per group, differential expression was defined as |FoldChange| > 2 and padj <0.05). (B) Volcano plot of DEGs highlighting genes related to sialic acid biosynthesis. (C) KEGG pathway enrichment analysis of upregulated DEGs in RB cells. (D,E) Immunofluorescence staining of α2,3-SA in mouse calvarial sections using MAL II lectin, with quantification of α2,3-SA fluorescence intensity (n = 5). Intensity density was normalized to the PBS group mean. (F) In vitro osteoclasts subjected to MAL II lectin staining and TRAP staining, with or without sialidase treatment to enzymatically remove α2,3-SA. (G) Quantification of α2,3-SA fluorescence intensity in cultured osteoclasts (n = 5). Intensity density was measured in cellular ROIs after background subtraction and normalized to the RANKL group mean. (H) Quantification of TRAP + multinucleated cells (≥3 nuclei) per field (randomly selected fields, fixed magnification) in vitro (n = 5). (I) mRNA expression levels of osteoclast differentiation markers ( Fos, Mmp9, Nfatc1, and Ocstamp ) in osteoclasts (n = 3). Data are presented as mean ± SD. Statistical significance was determined by two-tailed unpaired Student’s t-test for two-group comparisons (E) and one-way ANOVA followed by Tukey’s post hoc test for three-group comparisons (G–I) .

Article Snippet: Biotin MAL-II , Vector laboratories , Cat# B-1265-1.

Techniques: Activity Assay, Comparison, Infection, Quantitative Proteomics, Immunofluorescence, Staining, Fluorescence, In Vitro, Cell Culture, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Resolving heterogeneity of targeted lipid nanoparticles through solution-based biophysical analyses

doi: 10.64898/2026.03.31.715590

Figure Lengend Snippet: ( a ) SAXS profiles of NT LNPs and tLNPs, showing the characteristic Bragg peak feature associated with internal lipid–RNA organization. ( b ) Fitting of the Bragg peak feature using a multiple-Lorentz model where red is the first-order Bragg peak fit, green represents higher-order particle disorder and blue is the cumulative fit of the two features. (c) In-line AF4-UV-Vis contour maps (200–300 nm) depicting wavelength-resolved absorbance of LNPs during separation. The 260 nm absorbance signal is characteristic of encapsulated RNA, enabling identification of RNA-containing LNP populations across the AF4 elution profile. Overlapping spectral features indicate the presence of multiple co-eluting populations with distinct compositional profiles, motivating subsequent chemometric deconvolution. ( d ) Absorbance at 260 nm (RNA-associated signal) and 280 nm (protein-associated signal) from ( c ) plotted against the corresponding 260:280 ratio for each LNP formulation. Deviations in the 260:280 ratio across elution time indicate heterogeneity in RNA and protein content, suggesting the presence of compositionally distinct subpopulations that cannot be resolved by bulk measurements alone. These data were further subjected to chemometric analysis (see Supplemental Figure 6). The determined ( e ) R h profiles derived from in-line DLS and ( f ) molar mass profiles derived from MALS analysis for NT LNPs (beige) and tLNPs (colors) overlaid with UV fractograms from in-line AF4 separation. ( g ) Peak 260:280 ratios from ( d ), shown for comparison across LNP groups. ( h ) In-line DLS R h and MALS-derived ( i ) mass, ( j ) radius of gyration, and ( k ) polydispersity plotted for comparison across LNP groups. Measurements are reported mean ± standard error for ( h–k).

Article Snippet: A DAWN TM HELEOS II MALS instrument with an integrated WyattQELS TM DLS detector (Wyatt Technology), an Optilab TM T-rEX differential refractometer (Wyatt Technology), and a G7165A UV-Vis multi-wavelength UV-Vis detector (Agilent Technologies) were used for online detection.

Techniques: Formulation, Derivative Assay, Comparison

Journal: bioRxiv

Article Title: Resolving heterogeneity of targeted lipid nanoparticles through solution-based biophysical analyses

doi: 10.64898/2026.03.31.715590

Figure Lengend Snippet: ( a ) Guinier analyses with corresponding residuals for SVD-resolved C1 and C2 components of NT LNPs and tLNPs, with the exception of F(ab’) 2 tLNPs, where only C3 is shown. White regions indicate components for which Guinier analysis failed due to large size (q min R g > 1.3). ( b ) P(r) analyses normalized by I(0) for the average profile and individual components (C1–C3) for NT LNPs and tLNPs. ( c ) Radius of gyration (R g ) and ( d ) maximum dimension (D max ) of the average profile and individual components derived from GNOM analysis for NT LNPs and tLNPs. Blank regions denote populations where GNOM analysis was invalid (q min D max > 4). ( e ) LNP shape factor calculated as D max / R g , where values of ∼2.58 and ∼3.0 correspond to spherical and prolate ellipsoid geometries, respectively. DENSS ab initio electron density reconstructions from the AF4-UV-DLS-MALS-SAXS profiles for ( f ) NT LNPs, ( g ) nanobody tLNPs, ( h ) DAPRin tLNPs, ( i ), F(ab’) 2 tLNPs, and ( j ) antibody tLNPs.

Article Snippet: A DAWN TM HELEOS II MALS instrument with an integrated WyattQELS TM DLS detector (Wyatt Technology), an Optilab TM T-rEX differential refractometer (Wyatt Technology), and a G7165A UV-Vis multi-wavelength UV-Vis detector (Agilent Technologies) were used for online detection.

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Resolving heterogeneity of targeted lipid nanoparticles through solution-based biophysical analyses

doi: 10.64898/2026.03.31.715590

Figure Lengend Snippet: Targeted mRNA delivery to the placenta is driven by tLNP structural subspecies. ( a-d ) DiR-labeled NT LNPs and tLNPs containing mCherry mRNA were incubated with placental BeWo b30 trophoblasts at a dose of 150 ng of mRNA per 150,000 cells. After ( a ) 1 h, ( b ) 4 h, and ( c ) 24 h, cellular accumulation was quantified. After ( d ) 24 h, mCherry expression was also quantified. Normalized DiR and mCherry MFI was calculated by normalizing to cells treated with NT LNPs. ( e–m ) NT LNPs and tLNPs containing FLuc mRNA were administered intravenously via retroorbital injection into pregnant and nonpregnant mice at a dose of 12 µg mRNA per mouse. After 6 h, mice were euthanized, and major organs were dissected. For pregnant mice, luminescence imaging of ( e ) livers and spleens and ( f ) placentas and fetuses were performed via an in vivo imaging system (IVIS). Luminescence from ( e-f ) was quantified via region of interest (ROI) analysis to obtain luminescence flux in the ( g ) liver, ( h ) spleen, ( i ) placentas, and ( j ) fetuses of pregnant mice. For nonpregnant mice, luminescence imaging of ( k ) livers and spleens was performed. Luminescence from ( k ) was quantified via region of interest (ROI) analysis to obtain luminescence flux in the ( m ) liver and ( m ) spleen of nonpregnant mice. Signal is reported mean ± SD from n = 3 biological replicates for ( a–d ) and n = 4 biological replicates for ( e–m). One-way ANOVA with post hoc Student’s t-tests using the Holm–Sídak correction for multiple comparisons was used to compare fluorescence in for ( a–d ) and luminescence in ( g–h, l–m ) across treatment groups. Nested one-way ANOVA with post hoc Student’s t-tests using the Holm–Sídak correction for multiple comparisons was used to compare luminescence in ( i–j ) across treatment groups. ( n–q ) Spearman correlations for ( n ) placental, ( o ) pregnant hepatic, and ( p ) nonpregnant hepatic luminescence values using the physicochemical parameters from traditional characterization methods, static SAXS analyses, and AF4-UV-DLS-MALS-SAXS analyses. ( q ) Heatmap representing the entire dataset. For Spearman correlation graphs, dotted lines represent r = –0.6 and 0.6.

Article Snippet: A DAWN TM HELEOS II MALS instrument with an integrated WyattQELS TM DLS detector (Wyatt Technology), an Optilab TM T-rEX differential refractometer (Wyatt Technology), and a G7165A UV-Vis multi-wavelength UV-Vis detector (Agilent Technologies) were used for online detection.

Techniques: Labeling, Incubation, Expressing, Injection, Imaging, In Vivo Imaging, Fluorescence

Journal: bioRxiv

Article Title: Resolving heterogeneity of targeted lipid nanoparticles through solution-based biophysical analyses

doi: 10.64898/2026.03.31.715590

Figure Lengend Snippet: ( a-b ) NT LNPs and tLNPs containing FLuc mRNA were administered intravenously via retroorbital injection into pregnant and nonpregnant mice at a dose of 12 µg mRNA per mouse. After 6 h, mice were euthanized, and serum was collected. Serum levels of C3a, TNF, IFN-γ, IL-6, ALT, and AST were quantified in ( a ) pregnant and ( b ) nonpregnant mice via ELISA. Measurements are reported mean ± SD from n = 3–4 biological replicates. One-way ANOVA with post hoc Student’s t-tests using the Holm–Sídak correction for multiple comparisons was used to compare cytokine levels across treatment groups. ( c–e ) Spearman correlations for ( c ) TNF, ( d ) IFN-γ, and ( e ) IL-6 serum levels in pregnant (top) and nonpregnant (bottom) mice using the physicochemical parameters from traditional characterization methods, static SAXS analyses, and AF4-UV-DLS-MALS-SAXS analyses. ( f ) Heatmap representing the entire dataset. For Spearman correlation graphs, dotted lines represent r = –0.6 and 0.6.

Article Snippet: A DAWN TM HELEOS II MALS instrument with an integrated WyattQELS TM DLS detector (Wyatt Technology), an Optilab TM T-rEX differential refractometer (Wyatt Technology), and a G7165A UV-Vis multi-wavelength UV-Vis detector (Agilent Technologies) were used for online detection.

Techniques: Injection, Enzyme-linked Immunosorbent Assay