humalog  (Eli Lilly)

Journal: Journal of Diabetes Research

Article Title: Towards the Development of an Insulin Degradation Test

doi: 10.1155/jdr/8533644

Figure Lengend Snippet: Insulin absorbance readings do not detect insulin fibrillation. (A) Absorbance at 600 nm and (B) ThT fluorescence of Humalog, Novolog, and Basaglar after 96 h of exposure to each environmental stressor. However, the UV exposure was an accelerated stress condition and applied for only 2 h, as prolonged exposure caused rapid insulin deterioration and visible color change. Three replicates were used for each trial, and all trials were compared with 4°C using ANOVA with Dunnett correction. (C) Absorbance at 600 nm and (D) ThT fluorescence of unaltered Humalog insulin at ideal storage conditions of 4°C, Humalog at 65°C for 48 h, and Humalog at 65°C for 96 h. Three replicates were used for each trial, and each pairwise comparison was assessed using ANOVA with Tukey–Kramer correction. The asterisks denote ∗∗∗∗∗ p < 10 −5 , ∗∗∗∗ p < 10 −4 , ∗∗∗ p < 10 −3 , ∗∗ p < 10 −2 , and ∗ p < 5 ∗ 10 −2 .

Article Snippet: Experiments were conducted with new, unopened insulin vials and pens of 100 unit/mL Humalog (Eli Lilly), Novolog (Novo Nordisk), and Basaglar (Eli Lilly).

Techniques: Fluorescence, Comparison

Journal: Journal of Diabetes Research

Article Title: Towards the Development of an Insulin Degradation Test

doi: 10.1155/jdr/8533644

Figure Lengend Snippet: Insulin′s bioactivity decreases when fibrillation is above analog‐specific thresholds. (A) CHO cell bioactivity plotted against ThT fluorescence for Humalog degraded by various methods. For each scatter plot in this figure, the black curve is a power law function fit to the scatterplot data, with the p value and Pearson′s r value of the fit displayed. For all CHO experiments, at least three independent replicates were used (Methods) unless otherwise stated. (B) The CHO cell bioactivity and (C) ThT fluorescence of each degradation method for Humalog are shown. All trials in B and C were compared with 4°C using ANOVA with Dunnett correction. (D) CHO cell bioactivity plotted against ThT fluorescence for Novolog degraded by various methods. (E) The CHO cell bioactivity and (F) ThT fluorescence of each degradation method for Novolog are shown. All trials in E and F were compared with 4°C using ANOVA with Dunnett correction. (G) CHO cell bioactivity plotted against ThT fluorescence for Basaglar degraded by various methods. (H) The CHO cell bioactivity and (I) ThT fluorescence of each degradation method for Basaglar are shown. (J) Aliquots of one Basaglar vial were either kept at ideal conditions or were exposed to 37°C with agitation for 3, 3.5, 4, or 7 days. The 37°C with agitation for 3.5 days results were not statistically compared with the 4°C control in Panels H and I due to low independent replicates ( n = 2). All other trials in H, I, and J were compared with 4°C using ANOVA with Dunnett correction. (K) Two different vials of Basaglar were aliquoted and exposed to 37°C with agitation for 7 days. Three aliquots were tested from each vial and were compared by a two‐way t ‐test. The asterisks denote ∗∗∗∗∗ p < 10 −5 , ∗∗∗∗ p < 10 −4 , ∗∗∗ p < 10 −3 , ∗∗ p < 10 −2 , and ∗ p < 5 ∗ 10 −2 .

Article Snippet: Experiments were conducted with new, unopened insulin vials and pens of 100 unit/mL Humalog (Eli Lilly), Novolog (Novo Nordisk), and Basaglar (Eli Lilly).

Techniques: Fluorescence, Control

Journal: Journal of Diabetes Research

Article Title: Towards the Development of an Insulin Degradation Test

doi: 10.1155/jdr/8533644

Figure Lengend Snippet: Insulin′s secondary structure decreases α ‐helix signatures during fibrillation. (A) Far‐UV CD spectra were measured of fibrillated Humalog, Novolog, and Basaglar insulin samples that span the CHO bioactivity curves from Figure . The dots indicate each CD sample′s ThT measurement and corresponding projected bioactivity. (B) CD spectra of Humalog insulin exposed to ideal storage conditions of 4°C, 65°C for 48 h, and 65°C for 96 h. For all spectra in this Figure, the bold line is the mean of three replicates, and the shading around each mean spectrum is ± the standard deviation. The magnitude of (C) 208 nm, (D) 222 nm, and (E) 222 nm divided by 208 nm CD measurements are quantified. These wavelength measurements are indicative of protein secondary structure. (F) CD spectra of Novolog insulin exposed to ideal storage conditions of 4°C, 65°C for 48 h, and 65°C for 96 h. The magnitude of (G) 208 nm, (H) 222 nm, and (I) 222 nm divided by 208 nm measurements are quantified. (J) CD spectra of Basaglar insulin exposed to ideal storage conditions of 4°C, 37°C and agitation for 7 days, and 37°C for 14 days. The magnitude of (K) 208 nm, (L) 222 nm, and (M) 222 nm divided by 208 nm measurements are quantified. In the bar graphs, each pairwise comparison was assessed using ANOVA with Tukey–Kramer correction. The asterisks denote ∗∗∗∗∗ p < 10 −5 , ∗∗∗∗ p < 10 −4 , ∗∗∗ p < 10 −3 , ∗∗ p < 10 −2 , and ∗ p < 5 ∗ 10 −2 .

Article Snippet: Experiments were conducted with new, unopened insulin vials and pens of 100 unit/mL Humalog (Eli Lilly), Novolog (Novo Nordisk), and Basaglar (Eli Lilly).

Techniques: Circular Dichroism, Standard Deviation, Comparison

Journal: Journal of Diabetes Research

Article Title: Towards the Development of an Insulin Degradation Test

doi: 10.1155/jdr/8533644

Figure Lengend Snippet: Antibody panel can detect fibril proteins formed from thermal exposures. (A) ThT readings of the insulin used for the dot blot experimentation. Each pairwise comparison was assessed using ANOVA with Tukey–Kramer correction. The asterisks denote ∗∗∗∗∗ p < 10 −5 , ∗∗∗∗ p < 10 −4 , ∗∗∗ p < 10 −3 , ∗∗ p < 10 −2 , and ∗ p < 5 ∗ 10 −2 . (B) The ThT readings of the insulin analogs are denoted as dots on the CHO bioactivity curves from Figure , showing the samples′ projected bioactivities. Results of the fibril antibodies′ dot blots for (C) Humalog, (D) Novolog, and (E) Basaglar are shown in compiled images. Each antibody is denoted a different mOC number. A β 42 was the positive control, and TBS was the negative control. Each antibody per insulin type was a separate experiment and was cropped together to create Panels C–E. The orange dotted lines are where the images were cropped. The full dot blot membranes are shown in Figure .

Article Snippet: Experiments were conducted with new, unopened insulin vials and pens of 100 unit/mL Humalog (Eli Lilly), Novolog (Novo Nordisk), and Basaglar (Eli Lilly).

Techniques: Dot Blot, Comparison, Positive Control, Negative Control