Biology subjects

Haq, S. R.

Publications and source records attributed to Haq, S. R..

4 recordsLinked to original sources

bioRxiv2024

Preclinical study of the human recombinant Erythropoietin (GBPD002) compared with Eprex

Erythropoietin (EPO) is a glycoprotein that an essential growth factor for erythroid progenitors at the bone marrow, which appears in blood as a response to hypoxia. It is produced mainly by the kidneys; and its biosynthesis and release are stimulated by the reduction of tissue oxygenation and/or the reduction of the mass of erythrocytes. Here, we report the results of the preclinical evaluation of the safety of GBPD002 a recombinant human erythropoietin (rhEPO) developed by the Globe Biotech Limited, Bangladesh, through a comparative study of acute toxicity with Eprex(R), a commercial homologue from Janssen, UK. The product was administered subcutaneously into Wistar rats, at 500 lU/kg of therapeutic dose (TD) and 3 times of TD for the single dose toxicity study on 14 consecutive days and 125 UL/kg, 250 UL/kg, 500 UL/kg, 750 UL/kg, 1000 UL/kg, 1250 UL/kg and 1500 UL/kg of GBPD002 and Eprex(R) on 7 consecutive days respectively for the repeated dose toxicity study. Hematological and biochemical parameters were measured for all test subjects before first dose injection and the day after last dose injection of the both studies. Necropsy and histopathology of representative subjects from each group were also observed to find any pathological significance like degeneration or cellular necrosis in internal organs such as kidney, liver, lung and spleen of any rat under experiment. Both GBPD002 and Eprex(R) comparative toxicology studies revealed similar pharmacologically driven mechanisms of toxicity, which is statistically insignificant (p >0.05). Though hematology parameter values stayed within the normal range during the assay period but the high count of hemoglobin and high hematocrit (P<0.05), together with the decrease in white blood cell, confirm the therapeutic effect of Erythropoietin in both studies. Moreover, in both studies, the initial and final values of aspartate aminotransferase, alanine aminotransferase and blood urea nitrogen were also found similar (p >0.05) for both GBPD002 and Eprex(R) in the study. The study clearly established that the toxicological profile of "GBPD002" and Eprex(R), administered subcutaneously, were similar and related to the known pharmacology of erythropoietin alfa; hereby, demonstrating the proof of totality and no residual uncertainty between "GBPD002" and Eprex(R). Therefore, "GBPD002" and Eprex(R) shall be administered interchangeably in relevant indications.

pharmacology and toxicology↗

bioRxiv2023

Determining the shelf life of an erythropoietin alfa biosimilar GBPD002 through stability study

GBPD002 is a recombinant human erythropoietin (rhEPO) produced by recombinant DNA technology using mammalian cell expression system. In this study, samples were analyzed according to current Good Laboratory Practice (cGLP) and regulatory guidelines to evaluate the quality of the product under the influence of variety of environmental factors in a time-dependent manner. Accelerated (25 {+/-} 2 {degrees}C and relative humidity: 60 {+/-} 5 %) and real-time (5 {+/-} 3 {degrees}C) stability study were conducted up to 6 months and 12 months, respectively; samples were analyzed in every 3 months. After 12 months, real-time stability studies were performed with 6 months interval up to18 months, which to be continued up to 24 months. Appearance and pH were assessed using standard methods, and molecular weight was determined by Western blotting. Chromatographic identification and quantitative assays were performed by reversed- phase chromatography (RPC). High-molecular weight aggregatesand degradants were determined using size exclusion chromatography (SEC) and particle size distribution (PSD) analysis. Biofunctionality of the samples were evaluated by in vitro and in vivo bioassays. Bacterial endotoxin and sterility test were performed as safety parameters. All samples met the acceptance criteria, and the data were extrapolated using the regulatory guideline to determine the shelf life. The data revealed that the GBPD002 is stable up to 24 months.

pharmacology and toxicology↗

bioRxiv2020

Heimdallarchaea encodes profilin with eukaryotic-like actin regulation and polyproline binding

The evolutionary events which led to the first eukaryotic cell are still controversial1-4. The Asgard genome encodes a variety of eukaryotic signature proteins previously unseen in prokaryotes. Functional and structural characterization of these proteins is beginning to shed light on the complexity and pedigree of the ancestral eukaryotic cell5,6. In eukaryotes, the key cytoskeletal protein actin is important for diverse cellular processes such as membrane remodeling and cell motility7. Dynamic polymerization of actin provides both structure and generates the force which drives motility and membrane remodeling. These processes demand rapid filament assembly and disassembly on microsecond timescales. In eukaryotes, a variety of highly adapted proteins including gelsolin, profilin, VASP, ARP2/3 and signaling molecules (Phosphatidylinositol-4,5-bisphosphate (PIP2)) are crucial for organizing cellular cytoskeleton dynamics. Amongst others, the Asgard genomes encode predicted putative profilin homologues that regulate eukaryotic actin polymerization in vitro5,8. Interestingly, Asgard profilins appear to be regulated by PIP2, but not by polyproline rich motifs which are important for recruitment of actin:profilin complexes in eukaryotes5,9. These findings indicate that the Asgard archaea may have possessed analogous membrane organization to present-day eukaryotes, but that polyproline-mediated profilin regulation may have emerge later in the eukaryotic lineage5. Here, we show that Heimdallarchaeota, a candidate phylum within the Asgard superphylum, encodes a putative profilin (heimProfilin) that interacts with PIP2 and is regulated by polyproline motifs, implicative of an origin predating the rise of the eukaryotes. Additionally, we provide evidence for a novel regulatory mechanism whereby an extended N-terminal loop abolishes PIP2 and polyproline interactions. Lastly, we provide the first evidence for actin polymerization of an Asgard actin homologue. In context, these findings provide further evidence for the existence of a complex cytoskeleton already in Last eukaryotic common ancestor (LECA).

evolutionary biology↗

bioRxiv2020

NMR backbone assignment and dynamics of Profilin from Heimdallarchaeota

The origin of the eukaryotic cell is an unsettled scientific question. The Asgard superphylum has emerged as a compelling target for studying eukaryogenesis due to the previously unseen diversity of eukaryotic signature proteins. However, our knowledge about these proteins is still relegated to metagenomic data and very little is known about their structural properties. Additionally, it is still unclear if these proteins are functionally homologous to their eukaryotic counterparts. Here, we expressed, purified and structurally characterized profilin from Heimdallarchaeota in the Asgard superphylum. The structural analysis shows that while this profilin possess similar secondary structural elements as eukaryotic profilin, it contains additional secondary structural elements that could be critical for its function and an indication of divergent evolution.

biophysics↗