scholarly journals Recombinant Production of IL-1Ra in Fusion to Albumin Binding Domain for Its Extended Half-Life

2021 ◽  
Author(s):  
Fatemeh Shafiee ◽  
Ali Yazdani
Keyword(s):  
Mtt Assay ◽  
Half Life ◽  
Biological Effects ◽  
Binding Domain ◽  
Albumin Binding ◽  
Native Page ◽  
Biological Drug ◽  
E Coli ◽  
Recombinant Production ◽  
A375 Cells

Abstract Background: Anakinra, a FDA approved biological drug for Rheumatoid Arthritis, must be injected daily due to its short Half-life, leads to the lower patient compliance. So, the aim of this study was to produce IL-1Ra in fusing to albumin binding domain to extend its half-life and evaluate its biological effects.Methods and Results: The expression of IL-1Ra-ABD was performed in E. coli in fusing to intein1 of pTWIN1 in soluble and purified. The affinity of IL-1Ra-ABD to HSA was determined on Native-PAGE and its release percent toward time was determined. Finally, MTT assay was used to determine the antagonizing properties of recombinant IL-1Ra-ABD against IL-1β, on A375 cells. the expression induction of intein1-IL-1Ra-ABD using 0.1mM of IPTG at 15°C, and its cleavage represented a band approximately in 50 and 23 kDa respectively. Native-PAGE results showed that about 78% of IL-1Ra-ABD attached to the HSA after 2 hours of incubation, and MTT assay results showed no significant differences between the effects of our recombinant protein and native IL-1Ra.Conclusion: the production of soluble IL-1Ra-ABD with similar antagonizing effects to IL-1Ra was successfully performed. IL-1Ra-ABD showed suitable interaction with HSA and release over the time. However, pharmacokinetics and furthur biological evaluations are required.

A GLP-1 receptor agonist conjugated to an albumin-binding domain for extended half-life

Biopolymers ◽  
2014 ◽  
Vol 102 (3) ◽  
pp. 252-259 ◽  
Author(s):  
Joel Lindgren ◽  
Essam Refai ◽  
Sergei V. Zaitsev ◽  
Lars Abrahmsén ◽  
Per-Olof Berggren ◽  
...  
Keyword(s):  
Half Life ◽  
Receptor Agonist ◽  
Binding Domain ◽  
Albumin Binding

Fusion of an albumin-binding domain extends the half-life of immunotoxins

2016 ◽  
Vol 511 (1) ◽  
pp. 538-549 ◽  
Author(s):  
Rui Guo ◽  
Wenjun Guo ◽  
Li Cao ◽  
Hui Liu ◽  
Jieyu Liu ◽  
...  
Keyword(s):  
Half Life ◽  
Binding Domain ◽  
Albumin Binding

scholarly journals Extension of human GCSF serum half-life by the fusion of albumin binding domain

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Fatemeh Yadavar Nikravesh ◽  
Samira Shirkhani ◽  
Elham Bayat ◽  
Yeganeh Talebkhan ◽  
Esmat Mirabzadeh ◽  
...  
Keyword(s):  
Half Life ◽  
Animal Studies ◽  
Cytoplasmic Inclusion ◽  
Binding Domain ◽  
Size Exclusion ◽  
Albumin Binding ◽  
Exclusion Chromatography ◽  
Aggregation Formation

AbstractGranulocyte colony stimulating factor (GCSF) can decrease mortality of patients undergo chemotherapy through increasing neutrophil counts. Many strategies have been developed to improve its blood circulating time. Albumin binding domain (ABD) was genetically fused to N-terminal end of GCSF encoding sequence and expressed as cytoplasmic inclusion bodies within Escherichia coli. Biological activity of ABD-GCSF protein was assessed by proliferation assay on NFS-60 cells. Physicochemical properties were analyzed through size exclusion chromatography, circular dichroism, intrinsic fluorescence spectroscopy and dynamic light scattering. Pharmacodynamics and pharmacokinetic properties were also investigated in a neutropenic rat model. CD and IFS spectra revealed that ABD fusion to GCSF did not significantly affect the secondary and tertiary structures of the molecule. DLS and SEC results indicated the absence of aggregation formation. EC50 value of the ABD-GCSF in proliferation of NFS-60 cells was 75.76 pg/ml after 72 h in comparison with control GCSF molecules (Filgrastim: 73.1 pg/ml and PEG-Filgrastim: 44.6 pg/ml). Animal studies of ABD-GCSF represented improved serum half-life (9.3 ± 0.7 h) and consequently reduced renal clearance (16.1 ± 1.4 ml/h.kg) in comparison with Filgrastim (1.7 ± 0.1 h). Enhanced neutrophils count following administration of ABD-GCSF was comparable with Filgrastim and weaker than PEG-Filgrastim treated rats. In vitro and in vivo results suggested the ABD fusion as a potential approach for improving GCSF properties.

scholarly journals HER2-Specific Pseudomonas Exotoxin A PE25 Based Fusions: Influence of Targeting Domain on Target Binding, Toxicity, and In Vivo Biodistribution

Pharmaceutics ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 391
Author(s):  
Haozhong Ding ◽  
Mohamed Altai ◽  
Wen Yin ◽  
Sarah Lindbo ◽  
Hao Liu ◽  
...  
Keyword(s):  
Half Life ◽  
Binding Domain ◽  
Protein Domain ◽  
Specific Cell ◽  
Albumin Binding ◽  
Exotoxin A ◽  
Pseudomonas Exotoxin ◽  
Pseudomonas Exotoxin A ◽  
Number Of Patients ◽  
Plasma Half Life

The human epidermal growth factor receptor 2 (HER2) is a clinically validated target for cancer therapy, and targeted therapies are often used in regimens for patients with a high HER2 expression level. Despite the success of current drugs, a number of patients succumb to their disease, which motivates development of novel drugs with other modes of action. We have previously shown that an albumin binding domain-derived affinity protein with specific affinity for HER2, ADAPT6, can be used to deliver the highly cytotoxic protein domain PE25, a derivative of Pseudomonas exotoxin A, to HER2 overexpressing malignant cells, leading to potent and specific cell killing. In this study we expanded the investigation for an optimal targeting domain and constructed two fusion toxins where a HER2-binding affibody molecule, ZHER2:2891, or the dual-HER2-binding hybrid ZHER2:2891-ADAPT6 were used for cancer cell targeting. We found that both targeting domains conferred strong binding to HER2; both to the purified extracellular domain and to the HER2 overexpressing cell line SKOV3. This resulted in fusion toxins with high cytotoxic potency toward cell lines with high expression levels of HER2, with EC50 values between 10 and 100 pM. For extension of the plasma half-life, an albumin binding domain was also included. Intravenous injection of the fusion toxins into mice showed a profound influence of the targeting domain on biodistribution. Compared to previous results, with ADAPT6 as targeting domain, ZHER2:2891 gave rise to further extension of the plasma half-life and also shifted the clearance route of the fusion toxin from the liver to the kidneys. Collectively, the results show that the targeting domain has a major impact on uptake of PE25-based fusion toxins in different organs. The results also show that PE25-based fusion toxins with high affinity to HER2 do not necessarily increase the cytotoxicity beyond a certain point in affinity. In conclusion, ZHER2:2891 has the most favorable characteristics as targeting domain for PE25.

scholarly journals Albumin-binding domain extends half-life of glucagon-like peptide-1

2021 ◽  
Vol 890 ◽  
pp. 173650
Author(s):  
Huanbo Tan ◽  
Wencheng Su ◽  
Wenyu Zhang ◽  
Jie Zhang ◽  
Michael Sattler ◽  
...  
Keyword(s):  
Half Life ◽  
Binding Domain ◽  
Albumin Binding ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1

scholarly journals Preclinical Evaluation of Recombinant Human IL15 Protein Fused with Albumin Binding Domain on Anti-PD-L1 Immunotherapy Efficiency and Anti-Tumor Immunity in Colon Cancer and Melanoma

Cancers ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1789
Author(s):  
Fei-Ting Hsu ◽  
Yu-Chang Liu ◽  
Chang-Liang Tsai ◽  
Po-Fu Yueh ◽  
Chih-Hsien Chang ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Half Life ◽  
Significant Proportion ◽  
Suppressive Effect ◽  
Inhibitory Effect ◽  
Binding Domain ◽  
Albumin Binding ◽  
Effector Cells ◽  
Interleukin 15 ◽  
Immunosuppressive Cells

Anti-PD-L1 antibody monotherapy shows limited efficacy in a significant proportion of the patients. A common explanation for the inefficacy is a lack of anti-tumor effector cells in the tumor microenvironment (TME). Recombinant human interleukin-15 (hIL15), a potent immune stimulant, has been investigated in clinical trial with encouraging results. However, hIL15 is constrained by the short half-life of hIL15 and a relatively unfavorable pharmacokinetics profile. We developed a recombinant fusion IL15 protein composed of human IL15 (hIL15) and albumin binding domain (hIL15-ABD) and explored the therapeutic efficacy and immune regulation of hIL-15, hIL15-ABD and/or combination with anti-PD-L1 on CT26 murine colon cancer (CC) and B16-F10 murine melanoma models. We demonstrated that hIL15-ABD has significant inhibitory effect on the CT26 and B16-F10 tumor growths as compared to hIL-15. hIL-15-ABD not only showed superior half-life and pharmacokinetics data than hIL-15, but also enhance anti-tumor efficacy of antibody against PD-L1 via suppressive effect on accumulation of Tregs and MDSCs and activation of NK and CD8+T cells. Immune suppressive factors including VEGF and IDO were also decreased by combination treatment. hIL15-ABD combined with anti-PD-L1 antibody increased the activity of anti-tumor effector cells involved in both innate and adaptive immunities, decreased the TME’s immunosuppressive cells, and showed greater anti-tumor effect than that of either monotherapy.

scholarly journals Evaluation of an Albumin-Binding Domain Protein Fused to Recombinant Human IL-2 and Its Effects on the Bioactivity and Serum Half-Life of the Cytokine

2017 ◽  
Vol 21 (2) ◽  
pp. 77-83 ◽  
Author(s):  
Elham Adabi ◽  
Fateme Saebi ◽  
Amin Moradi Hasan-Abad ◽  
Ladan Teimoori-Toolabi ◽  
Gholam Ali Kardar ◽  
...  
Keyword(s):  
Half Life ◽  
Binding Domain ◽  
Albumin Binding ◽  
Domain Protein
Sign in / Sign up

Export Citation Format

Share Document