Hamid Bashir

Background: A major challenge in modern oncology is developing therapeutics that selectively eliminate tumor cells while sparing healthy tissues. Targeted delivery using engineered chimeric biomolecules can substantially augment therapeutic efficacy and safety. Interleukin-29 (IL-29/IFN-λ1) is a potent type III interferon with antitumor and immunomodulatory properties, whereas GW-H1 is a synthetic cationic antimicrobial peptide that induces caspase-dependent apoptosis in hepatocellular carcinoma (HCC) cells. In this study, an IL-29–GW-H1 chimeric protein was computationally designed by fusing human IL-29 to the GW-H1 peptide via a rigid helical linker [A(EAAAK)5A], followed by recombinant bacterial expression, purification, and in vitro functional validation. Methods: Primary amino acid sequences were retrieved and analyzed for physicochemical parameters, secondary structure, and three-dimensional (3D) folding using ProtParam, GOR IV, trRosetta, I-TASSER, and AlphaFold2. Structural stereochemistry was validated using SAVES v6.0 (PROCHECK, ERRAT, Verify3D) and ProSA-web. Molecular docking was performed using ClusPro 2.0 against the IL-28Rα/IL-10Rβ heterodimeric receptor, followed by 100 ns molecular dynamics (MD) simulations in Desmond (Schrödinger) to analyze conformational stability, root mean square deviation (RMSD), root mean square fluctuation (RMSF), dynamic cross-correlation, and binding free energy via MM/GBSA. The chimeric gene was synthesized, cloned into the pET29b(+) expression vector, expressed in Escherichia coli BL21(DE3), isolated from inclusion bodies, refolded, and purified via Ni-NTA affinity chromatography. In vitro cytotoxicity was evaluated against HepG2 hepatoma cells and non-tumorigenic HEK293 cells using the MTT assay. Results: The refined 3D model exhibited excellent structural quality, with an ERRAT score of 97.06% and 98% of residues situated within the most favored regions of the Ramachandran plot. Molecular docking demonstrated favorable binding of the chimeric construct to the IL-28Rα/IL-10Rβ receptor complex, mediated by 8 salt bridges, 28 hydrogen bonds, and a favorable binding free energy (ΔG = −116.31 kcal/mol). MD simulations over 100 ns confirmed structural equilibration, minimal backbone fluctuations, and a well-defined global free energy minimum. Recombinant IL-29–GW-H1 was successfully expressed in E. coli and purified to >98% purity (final yield: 63 mg/L). In MTT cytotoxicity assays, the chimeric protein exhibited significantly higher cytotoxic potency against HepG2 cells (IC50 = 16.31 µg/mL) than wild-type IL-29 (IC50 = 20.14 µg/mL; p < 0.001), while displaying minimal cytotoxicity toward normal HEK293 cells (>60% viability at 25 µg/mL). Conclusion: The engineered IL-29–GW-H1 fusion protein demonstrates robust structural stability, strong receptor affinity, and enhanced tumor-selective cytotoxicity against hepatocellular carcinoma cells in vitro. These findings suggest that this novel chimeric construct represents a promising targeted candidate for liver cancer immunotherapy.

Introduction: Targeted cancer therapy using recombinant fusion proteins offers a novel and precise approach to addressing limitations associated with conventional treatments, such as nonspecific toxicity and drug resistance. Fusion proteins are designed to synergistically incorporate multiple protein domains, thereby enhancing bioactivity and creating novel functional combinations with broad applications. In this study, we report the innovative design and evaluation of a fusion protein that combines interleukin-24 (IL-24), a tumor-suppressive cytokine, with melittin, a potent cell-lytic peptide derived from honey-bee venom. Methods: This combination synergistically unites the pro-apoptotic properties of IL-24 with the membrane-disrupting activity of melittin to increase cytotoxicity against cancer cells. The Melittin-IL-24 construct was expressed in Escherichia coli BL21(DE3) and induced with IPTG. Results: After affinity chromatography, we obtained ~100 mg of > 97 %-pure fusion protein per liter of culture, as verified by SDS-PAGE, Western blotting, and HPLC. MTT assays showed significant cytotoxicity of Melittin-IL-24 toward MCF-7 breast-cancer cells (IC₅₀ = 52 µg/mL), whereas IL-24 alone exhibited an IC₅₀ of 66 µg/mL. Both proteins were far less toxic to normal MCF-10F mammary epithelial cells. Conclusion: By combining the apoptotic effects of IL-24 with the cell-lytic properties of melittin, Melittin-IL-24 demonstrates enhanced therapeutic potential and may represent a promising strategy for targeted cancer therapy.

Tumor targeting peptides (TTPs) have emerged as new therapeutic and diagnostic tools in oncology, due to their low immunogenicity, high specificity, and ability to efficiently penetrate tumor cells and tissues. They exert their effects using various mechanisms such as receptor-mediated targeting, cell-penetrating properties, and enzyme-responsive activation, allowing selective delivery of drugs, nanoparticles, and imaging agents to cancer cells. Advances in peptide engineering, such as D-amino acid incorporation, cyclization, and multivalent designs, have substantially enhanced their stability, affinity, and bioavailability. They are widely utilized in immunotherapy, precision imaging, and targeted drug delivery, thus improving cancer detection and outcomes. Recent developments, including peptide–drug conjugates, hybrid peptide–nanoparticle systems, and peptide-based immune modulators, have significantly broadened the clinical potential of TTPs. This review highlights the fundamental mechanisms, therapeutic applications, and cutting-edge advancements in TTPs, underscoring their role in personalized cancer therapy.

Mycoplasma genitalium is increasingly recognized for its role in severe health conditions, including sexually transmitted infections, ovarian and prostate cancer. The adhesion protein plays a crucial role in the pathogen’s ability to attach to and invade host cells, making it a key target for vaccine development. The need to develop a vaccine against M. genitalium stems from its rising antibiotic resistance, limited treatments and effectiveness. This study focuses on the design and computational evaluation of adhesion protein-based epitope vaccine. Through an immunoinformatic approach, multiple novel cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL), and linear B-cell epitopes were identified from the adhesion protein, demonstrating strong antigenic, non-allergenic, and immunogenic properties. The vaccine construct’s 3D structure was validated using Ramachandran plot analysis, ProSA, and ERRAT servers, confirming its stability and suitability. Molecular docking studies revealed a high binding affinity of the vaccine with the TLR-2 receptor, further supported by 100 ns molecular dynamics (MD) simulations that confirmed the structural stability and robust interaction of the vaccine with immune receptors. In silico immune simulations using the C-ImmSim server demonstrated the vaccine’s potential to elicit strong humoral and cell-mediated immune responses. Codon optimization for expression in E. coli using the pET-29a(+) vector predicted efficient production of the vaccine. The comprehensive computational analysis, underscores the potential of this epitope-based vaccine as a promising candidate against M. genitalium infections. However, the study emphasizes the necessity of in vitro and in vivo experiments to validate the vaccine’s efficacy and safety before advancing to clinical trials.

Introduction: Synthetic fusion proteins represent a cutting-edge approach in biotechnology and pharmaceutical research, enabling the strategic combination of multiple protein domains to design novel complexes with enhanced properties and functionalities. By fusing distinct proteins, the unique attributes of each component can be synergistically exploited, leading to improved bioactivities or the emergence of entirely new functions. This study aimed to computationally construct a fusion protein that contained the killing properties of temporin 1CEa and the targeting action of IL-24, joined via a rigid linker, and its anti-tumor potential was analyzed using different bioinformatics tools. Methods: After sequence retrieval, the 3D model of the temporin 1CEa-IL-24 fusion protein was constructed using the AlphaFold 2 online server. The resulting structure underwent rigorous refinement, quality assessment, and validation processes. Physiochemical properties were evaluated using ProtParam, and the prepared structure was subjected to docking on ClusPro and simulation on VMD and NAMD. Results: Assessment through ERRAT score and Ramachandran plot analysis confirmed the good quality, refinement, and validation of the 3D structure of the temporin 1CEa-IL-24 fusion protein. Docking results revealed 17 hydrogen bonds and 4 salt bridges between the fusion protein and its receptor, findings supported and validated through simulation studies predicting a stable docked complex over a 100 ns period. Conclusion: The expression of the temporin 1CEa-IL-24 fusion gene in a suitable expression host could lead to in vitro production and subsequently validate its therapeutic potential.

Targeted cancer therapy has emerged as a promising alternative to conventional chemotherapy, which is often plagued by poor selectivity, off-target effects, and drug resistance. Among the various targeting agents in developme... | Find, read and cite all the research you need on Tech Science Press

The immune system comprises various regulators and effectors that elicit immune responses against various attacks on the body. The pathogenesis of autoimmune diseases is derived from the deregulated expression of cytokines, the major regulators of the immune system. Among cytokines, interleukins have a major influence on immune-mediated diseases. These interleukins initiate the immune response against healthy and normal cells of the body, resulting in immune-mediated disease. The major interleukins in this respect are IL-1, IL-3, IL-4, IL-6, IL-10 and IL-12 which cause immune responses such as excessive inflammation, loss of immune tolerance, altered T-cell differentiation, immune suppression dysfunction, and inflammatory cell recruitment. Systemic Lupus Erythematosus (SLE) is an autoimmune illness characterized by dysregulation of interleukins. These immune responses are the signs of diseases such as rheumatoid arthritis, inflammatory bowel disease, psoriasis, type I diabetes, and multiple sclerosis.