Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Construction, purification, and characterization of FGF21-164. ( A ) FGF21-164 sequence and FGF21 sequence. In the FGF21-164 sequence, red marked the mutated amino acid and blue represents the TR acid sequence from hCD164. In the FGF21 sequence, the red-labeled amino acid represents the amino acid sequence whose n-terminal is deleted in FGF21-164. ( B ) Subcloned cell culture supernatants expressing FGF21-164 and FGF21 were analyzed using Western blotting (Lane M: molecular weight marker. Lane 1~7: FGF21-164 clones 1, 2, 3, A4, B2, F3, and H4. Line 8: FGF21 clone2). ( C ) The purified FGF21-164 and FGF21 were assayed using SDS-PAGE and Western blotting (Lane M: molecular weight marker. Lane 1: purified FGF21-164. Line 2: purified FGF21). ( D ) Purity and hydrodynamic radius of FGF21-164 were analyzed using SEC-HPLC. ( E ) MALDI-TOF mass-spectrometry analysis of FGF21-164. ([M + H] + denotes the singly charged ionic forms, and [M + 2H] 2+ denotes doubly charged ionic forms). ( F ) Circular Dichroism (CD) spectra of FGF21-164 and FGF21.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques: Purification, Sequencing, Labeling, Cell Culture, Expressing, Western Blot, Molecular Weight, Marker, Clone Assay, SDS Page, Mass Spectrometry, Circular Dichroism

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Glycan forms analysis of the protein FGF21-164. ( A ) O-linked glycan forms of FGF21-164. ( B ) N-linked glycan forms of FGF21-164.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of FGF21-164 in adipocytes induced by 3T3-L1. ( A ) The process of inducing 3T3-L1’s differentiation into adipocytes. ( B ) Reduction in glucose in medium stimulated by FGF21-164 in adipocytes induced by 3T3-L1. Data are means ± SEM (n = 6). * p < 0.05, *** p < 0.001, and **** p < 0.0001 compared to 0 μg/mL FGF21-164. ( C ) Phospho-Erk1/2-specific bands (Thr202/Tyr204; 44–42 kDa) were detected in 3T3-L1-derived adipocytes after FGF21-164 stimulation. ( D ) Evaluation of lipid droplet accumulation in adipocytes induced by 3T3-L1 with or without FGF21-164. The scale bar represents 50 μm.Data are the means ± SEM (n = 6). * p < 0.05, **** p < 0.0001.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Pharmacokinetics of FGF21-164. FGF21-164 plasma levels in C57BL/6 mice as determined via targeted LC-MS after i.v. (tail vein) bolus injection of FGF21-164 protein.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques: Liquid Chromatography with Mass Spectroscopy, Injection

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: The results of an oral glucose tolerance test applied to ob / ob mice after a single FGF21-164 treatment. ( A ) Blood glucose levels were measured at 0, 2, 3, 4, 5, 6, 7, and 8 h after the administration of FGF21-164. Data are the means ± SEM (n = 4–5). * p < 0.05 and ** p < 0.01 compared with the model. ( B ) The decrease in blood glucose levels post-administration relative to the baseline measurement at 0 h. Data are the means ± SEM (n = 4–5). * p < 0.05 compared with the model.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of a single administration of FGF21-164 on ob / ob mice. ( A , D , G ) Fasting glucose levels. Blood glucose levels were measured at 7, 21, and 28 days after a single dose of FGF21-164 was administered. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, ** p < 0.01, and **** p < 0.0001. ( B , E , H ) The OGTT at 7, 21, and 28 days. After oral glucose administration, blood glucose levels were measured. Data are the means ± SEM (n = 4–5). * p < 0.05, *** p < 0.001, and **** p < 0.0001 at 6 mg·kg −1 vs. model. # p < 0.05, ## p < 0.01 and #### p < 0.0001 at 12 mg·kg −1 vs. model. ( C , F , I ) The glucose AUC during the OGTT process. Data are the means ± SEM (n = 4–5). ns: no significance. * p < 0.05, *** p < 0.001. ( J ) Body weight measured at 0, 7, 21, and 28 days. Data are the means ± SEM (n = 4–5). ns: no significance.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: A Novel Recombinant Human FGF21 Analog with High Glycosylation Has a Prolonged Half-Life and Affects Glycemic and Body Weight Control

doi: 10.3390/ijms26062672

Figure Lengend Snippet: Effect of repeated administration of FGF21-164. ( A ) Body weight changes. The DIO mice were treated with PBS or FGF21-164 once every 2 days. Data are the means ± SEM (n = 5–7). **** p < 0.0001 vs. Ctrl. #### p < 0.0001 for FGF21-164 vs. the model. ( B ) The liver sections were stained with Oil-red-O. The scale bar represents 100 μm. DIO mice showed many red lipid droplets in hepatic tissue (indicated by the black arrow), while lipid droplet levels were lower after the FGF21-164 treatment.

Article Snippet: Recombinant protein expression was initially characterized using Western blot analysis via an anti-FGF21 antibody (#50421-R005, SinoBiologicals, Beijing, China), followed by protein fermentation and purification.

Techniques: Staining

Journal: Veterinary sciences

Article Title: Establishment of a Direct Competitive ELISA for Camel FGF21 Detection.

doi: 10.3390/vetsci12020170

Figure Lengend Snippet: Figure 1. Conservation analysis of FGF21 protein. (A) Sequence of camel, human, mouse, and pig FGF21 peptides were aligned with CLUSTALW. (B) Spatial structures of camel, human, mouse, and pig FGF21 protein were predicted with SWISS-MODEL, respectively.

Article Snippet: After blocking with 10% skimmed milk in TBST for 2 h at room temperature, the membranes were incubated overnight at 4 ◦C with primary antibodies, including the anti-camel FGF21 polyclonal antibody mentioned above, anti-His tag antibody (CusaBio, Wuhan, China), or anti-human FGF21 antibody (ab171941, Abcam, Vet.

Techniques: Sequencing

Journal: Veterinary sciences

Article Title: Establishment of a Direct Competitive ELISA for Camel FGF21 Detection.

doi: 10.3390/vetsci12020170

Figure Lengend Snippet: Figure 3. Purification and analysis of guinea pig anti-camel FGF21 polyclonal antibody. (A) UV profile during purification of guinea pig IgG using Protein A HP affinity chromatography. (B) SDS-PAGE analysis of purified guinea pig IgG. M: Protein marker. 1: Purified guinea pig IgG. (C) Antibody titer was determined by indirect ELISA mediated by camel FGF21-encapsulated antigen and HRP-labeled goat anti-guinea-pig IgG. (D) Antibody specificity was monitored using western blot. M: Protein marker. 1: Camel FGF21 protein. 2: Mouse FGF21 protein. 3: Human FGF21 protein. 4: Pig FGF21 protein (Supplementary File S1).

Article Snippet: After blocking with 10% skimmed milk in TBST for 2 h at room temperature, the membranes were incubated overnight at 4 ◦C with primary antibodies, including the anti-camel FGF21 polyclonal antibody mentioned above, anti-His tag antibody (CusaBio, Wuhan, China), or anti-human FGF21 antibody (ab171941, Abcam, Vet.

Techniques: Purification, Affinity Chromatography, SDS Page, Marker, Indirect ELISA, Labeling, Western Blot

Journal: Veterinary sciences

Article Title: Establishment of a Direct Competitive ELISA for Camel FGF21 Detection.

doi: 10.3390/vetsci12020170

Figure Lengend Snippet: Figure 6. Establishment of camel FGF21 detection standard curve.

Article Snippet: After blocking with 10% skimmed milk in TBST for 2 h at room temperature, the membranes were incubated overnight at 4 ◦C with primary antibodies, including the anti-camel FGF21 polyclonal antibody mentioned above, anti-His tag antibody (CusaBio, Wuhan, China), or anti-human FGF21 antibody (ab171941, Abcam, Vet.

Techniques:

Journal: Veterinary sciences

Article Title: Establishment of a Direct Competitive ELISA for Camel FGF21 Detection.

doi: 10.3390/vetsci12020170

Figure Lengend Snippet: Figure 7. Establishment of mouse, human and pig FGF21 detection standard curve.

Article Snippet: After blocking with 10% skimmed milk in TBST for 2 h at room temperature, the membranes were incubated overnight at 4 ◦C with primary antibodies, including the anti-camel FGF21 polyclonal antibody mentioned above, anti-His tag antibody (CusaBio, Wuhan, China), or anti-human FGF21 antibody (ab171941, Abcam, Vet.

Techniques: