Recombinant production of interleukin-1 receptor antagonist in fusion to albumin binding domain with potential affinity to human serum albumin

Fatemeh Shafiee , Ali Yazdani

Abstract


Background and purpose: Anakinra must be injected daily due to its short half-life and this leads to lower patient compliance. Therefore, the aim of this study was to produce an interleukin-1 receptor antagonist (IL-1Ra) with albumin binding domain (ABD) as a novel fusion protein and evaluate its binding ability to albumin and its biological effects.

Experimental approach: The three-dimensional structure of IL-1Ra-ABD was predicted by MODELLER software and its interaction with IL-1R was evaluated by the HADDOCK server. The expression of IL-1Ra-ABD was performed in E. coli in fusion with intein 1 of pTWIN1 in soluble form and then purified. The affinity of IL-1Ra-ABD to human serum albumin (HSA) was determined on native-PAGE, and its release percent toward time was evaluated. Moreover, an MTT assay was used to determine the antagonizing properties of recombinant IL-1Ra-ABD against IL-1β in A375 and HEK293 cell lines.

Findings/Results: The stable complex of IL-1Ra-ABD with IL-1R established the absence of steric hindrance due to the addition of ABD to IL-1Ra. The expression induction of intein 1-IL-1Ra-ABD using 0.1 mM IPTG at 15 °C, and its cleavage represented bands approximately in 50 and 23 kDa. Furthermore, about 78% of IL-1Ra-ABD was attached to the HSA after 2 h of incubation, and the MTT assay showed no significant differences between the effects of IL-1Ra-ABD and native IL-1Ra in cell survival.

Conclusions and implications: The production of soluble IL-1Ra-ABD with no significant differences in IL-1Ra antagonizing effects was successfully performed. IL-1Ra-ABD showed suitable interaction with HSA and was released over time. However, the half-life of IL-1Ra-ABD in vivo must be determined in the subsequent investigations.


Keywords


Albumin binding domain; Human serum albumin; IL-1Ra; Intein.

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References


Schnellbacher C, Ciocca G, Menendez R, Aksentijevich I, Goldbach‐Mansky R, Duarte AM, et al. Deficiency of interleukin‐1 receptor antagonist responsive to anakinra. Pediatr Dermatol. 2013;30(6):758-760.DOI: 10.1111/j.1525-1470.2012.01725x.

Waugh J, Perry CM. Anakinra. BioDrugs. 2005;19(3):189-202.DOI: 10.2165/00063030-200519030-00005.

Kaiser C, Knight A, Nordström D, Pettersson T, Fransson J, Florin-Robertsson E, et al. Injection-site reactions upon kineret (anakinra) administration: experiences and explanations. Rheumatol Int. 2012;32(2):295-299. DOI: 10.1007/s00296-011-2096-3.

Suk JS, Xu Q, Kim N, Hanes J, Ensign LM. PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev. 2016;99(Pt. A):28-51. DOI: 10.1016/j.addr.2015.09.012.

Liebner R, Mathaes R, Meyer M, Hey T, Winter G, Besheer A. Protein HESylation for half-life extension: synthesis, characterization and pharmacokinetics of HESylated anakinra. Eur J Pharma Biopharm. 2014;87(2):378-385.DOI: 10.1016/j.ejpb.2014.03.010.

Podust VN, Balan S, Sim BC, Coyle MP, Ernst U, Perers RT, et al. Extension of in vivo half-life of biologically active molecules by XTEN protein polymers. J Control Release. 2016; 240:52-66. DOI: 10.1016/j.jconrel.2015.10.038.

Bern M, Nilsen J, Ferrarese M, Sand KMK, Gjølberg TT, Lode HE, et al. An engineered human albumin enhances half-life and transmucosal delivery when fused to protein-based biologics. Sci Transl Med. 2020;12(565):eabb0580,1-13.DOI: 10.1126/scitranslmed.abb0580.

Liu M, Huang Y, Lei Hu, Liu G, Hu X, Liu D, et al. Selective delivery of interleukine-1 receptor antagonist to inflamed joint by albumin fusion. BMC Biotechnol. 2012;12:68,1-13.DOI: 10.1186/1472-6750-12-68.

Mester S, Evers M, Meyer S, Nilsen J, Greiff V, Sandlie I, et al. Extended plasma half-life of albumin-binding domain fused human IgA upon pH-dependent albumin engagement of human FcRn in vitro and in vivo. MAbs. 2021;13(1):e1893888,1-13. DOI: 10.1080/19420862.2021.1893888.

Levy OE, Jodka CM, Ren SS, Mamedova L, Sharma A, Samant M, et al. Novel exenatide analogs with peptidic albumin binding domains: potent anti-diabetic agents with extended duration of action. PLoS One. 2014;9(2):e87704,1-9.DOI: 10.1371/journal.pone.0087704.

Sheffield WP, Smith IJ, Syed S, Bhakta V. Prolonged in vivo anticoagulant activity of a hirudin-albumin fusion protein secreted from Pichia pastoris. Blood Coagul Fibrinolysis. 2001;12(6):433-443.DOI: 10.1097/00001721-200109000-00003.

Lindgren J, Refai E, Zaitsev SV, Abrahmsén L, Berggren PO, Karlström AE. A GLP-1 receptor agonist conjugated to an albumin-binding domain for extended half-life. Biopolymers. 2014;102(3):252-259. DOI: 0.1002/bip.22474.

Jacobs SA, Gibbs AC, Conk M, Yi F, Maguire D, Kane C, et al. Fusion to a highly stable consensus albumin binding domain allows for tunable pharmacokinetics. Protein Eng Des Sel. 2015;28(10):385-393. DOI: 10.1093/protein/gzv040.

Barkestani MN, Naserian S, Khoddam F, Shamdanu S, Bambai B. Optimization of IL-1RA structure to achieve a smaller protein with a higher affinity to its receptor. Sci Rep. 2022;12(1):7483,1-9.DOI: 10.1038/s41598-022-11100-3.

Adelnia R, Shafiee F. Recombinant production and one step purification of il-1ra in Escherichia coli and evaluation its il-1 antagonizing efficacy. Iran J Immunol. 2021;18(2):141-149.DOI: 10.22034/iji.2021.89103.1929.

Sima S, Shafiee F, Jahanian-Najafabadi A. Expression and one step intein-mediated purification of biologically active human G-CSF in Escherichia coli. Mol Biol Rep. 2020;47(4):2861-2869.DOI: 10.1007/s11033-020-05404-8.

Jiang A, Jin W, Zhao F, Tang Y, Sun Z, Liu JN. Split Ssp DnaB mini-intein-mediated production of recombinant human glucagon-like peptide-1/7-36. Biotechnol Appl Biochem. 2015;62(3):309-315. DOI: 10.1002/bab.1274.

Zhang M, Zhang Y, Wu B, Peng Y, Simair AA, Siegel GW, et al. Intein-mediated recombinant expression of monomeric B22Asp desB30 insulin. BMC Biotechnol. 2020; 20(1):3, 1-9. DOI: 10.1186/s12896-020-0598-3.

Wang C, Wang MW, Tashiro SI, Onodera S, Ikejima T. IL-1β acts in synergy with endogenous IL-1β in A375-S2 human melanoma cell apoptosis through mitochondrial pathway. J Korean Med Sci.

;20(4):555-561.DOI: 10.3346/jkms.2005.20.4.555.

Liu M, Huang Y, Hu L, Liu G, Hu X, Liu D, et al. Selective delivery of interleukine-1 receptor antagonist to inflamed joint by albumin fusion. BMC Biotechnol. 2012;12(1):68,1-13.DOI: 10.1186/1472-6750-12-68.

Wang YX, Yang ZX, Zhu HQ, Zhou XW, Huang PT. Construction, expression and preliminary pharmacokinetics of IL-1ra mutants. Chin J Biotechnol. 2006;22(3): 472-476.DOI: 10.1016/S1872-2075(06)60040-X.

Powers NE, Swartzwelter B, Marchetti C, de Graaf DM, Lerchner A, Schlapschy M, et al. PASylation of IL-1 receptor antagonist (IL-1Ra) retains IL-1 blockade and extends its duration in mouse urate crystal-induced peritonitis. J Biol Chem. 2020;295(3): 868-882.DOI: 10.1074/jbc.RA119.010340


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