Sharif Digital Repository

Background: Chinese hamster ovary (CHO) cell line is considered as the most common cell line in the biopharmaceutical industry because of its capability in performing efficient post-translational modifications and producing the recombinant proteins, which are similar to natural human proteins. The optimization of the upstream process via different feed strategies has a great impact on the target molecule expression and yield. Methods: To determine and understand the molecular events beneath the feed effects on the CHO cell, a label-free quantitative proteomic analysis was applied. The proteome changes followed by the addition of a designed amino acid feed to the monoclonal antibody producing... 

Artificial Immune System algorithms use antibodies which fully specify the solution of an optimization, learning, or pattern recognition problem. By being restricted to fully specified antibodies, an AIS algorithm can not make use of schemata or classes of partial solutions. This paper presents a symbiotic artificial immune system (SymbAIS) algorithm which is an extension of CLONALG algorithm. It uses partially specified antibodies and gradually builds up building blocks of suitable sub-antibodies. The algorithm is compared with CLONALG on multimodal function optimization and combinatorial optimization problems and it is shown that it can solve problems that CLONALG is unable to solve.... 

In the present study, the feasibility of electrospun polyethersolfone (PES) nanofibrous membrane as the solid substrate for microfluidic based immunoassays to enhance the density of immobilized antibody on the surface of membrane was assessed. Conversely, the efficacy of antibody immobilization was compared by two different strategies as 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)/N-Hydroxysuccinimide (NHS) coupling chemistry and hydrophobic interaction. Compared to conventional immunoassays carried out in plates or gels, microfluidic based immunoassays grant a lot of advantages such as a consumption of little samples and reagents, shorter analysis time, and higher efficiency.... 

In this paper we introduce novel strategy for antibody immobilization using high surface area electrospun nanofibrous membrane based on ethyl-3-(3-dimethylaminopropyl)-carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling chemistry. To present the high performance of proposed biosensors, anti-staphylococcus enterotoxin B (anti-SEB) was used as a model to demonstrate the utility of our proposed system. Polymer solution of polyethersolfone was used to fabricate fine nanofibrous membrane. Moreover, industrial polyvinylidene fluoride membrane and conventional microtiter plate were also used to compare the efficiency of antibody immobilization. Scanning electron microscopy images were taken to... 

The present study is aimed at the development of a novel approach based on the magnetic improvement of DNA-directed antibody immobilization to prepare a highly efficient sensing platform. Magnetic nanoparticle substrates with high surface area capture the dual DNA-conjugated antibodies in a solution. This allows overcoming the typical mass transport limitation of the surface-based antibody immobilization. Antibody-magnetic nanoparticle conjugation is based on a robust hybridization between a DNA tether (attached to the antibody) and its complementary sequence (immobilized on the nanoparticle). Conventional antibody immobilization for the detection of proteins is often insignificant for the... 

The present study is aimed at the synthesis of MnFe2O4@SiO2@PMIDA in terms of highly efficient sensing platform for anti-prostate specific membrane antigen (PSMA) immobilization. Superparamagnetic manganese ferrite nanoparticles were synthesized following co-precipitation method and then SiO2 shell was coated on the magnetic core with tetraethyl orthosilicate (TEOS) through a silanization reaction to prevent oxidation, agglomeration and, increase the density of OH groups on the surface of MnFe2O4. Subsequently, MnFe2O4@SiO2@PMIDA obtained as a result of the reaction between N-(phosphonomethyl)iminodiacetic acid (PMIDA) and MnFe2O4@SiO2. The reactive carboxyl groups on the surface of magnetic... 

Since physiological and pathological processes occur at nano-environments, nanotechnology has considered as an efficient tool for designing of next generation specific biomolecules with enhanced pharmacodynamic and pharmacodynamic properties. In the current investigation, by control of the size and hydrodynamic volume at the nanoscale, for the first time, physicochemical and pharmacokinetic properties of an anti-VEGFA nanobody was remarkably improved by attachment of a Proline-Alanine-Serine (PAS) rich sequence. The results elucidated unexpected impressive effects of PAS sequence on physicochemical properties especially on size, hydrodynamics radius, and even solubility of nanobody. CD... 

Celiac disease is an autoimmune disorder represented by the ingestion of the gluten protein usually found in wheat, barley and rye. To date, ELISA has been the most accurate method for determining the presence of anti-gliadin, which is cumbersome, expensive (compared to a suspension microarray technique), and requires extensive sample preparation. In this study, in order to establish a more accurate assay to identify gliadin at lower concentrations, optical nano biosensors using an indirect immunoassay method for gliadin detection was designed and fabricated. For this, polycaprolactone (PCL) nano- to micro-beads were fabricated as a platform for the gliadin antigen which were optimized and... 

Coronaviruses (CoV) are a family of viral pathogens that infect both birds and mammals, including humans. Seven human coronaviruses (HCoV) have been recognized so far. HCoV-229E, -OC43, -NL63, and -HKU1 account for one-third of common colds with mild symptoms. The other three members are severe acute respiratory syndrome (SARS)-CoV, Middle East respiratory syndrome (MERS)-CoV, and SARS-CoV-2. These viruses are responsible for SARS, MERS, and CoV disease 2019 (COVID-19), respectively. A variety of diagnostic techniques, including chest X-rays, computer tomography (CT) scans, analysis of viral nucleic acids, proteins, or whole virions, and host antibody detection using serological assays have... 

Cell culture feeds optimization is a critical step in process development of pharmaceutical recombinant protein production. Amino acids are the basic supplements of mammalian cell culture feeds with known effect on their growth promotion and productivity. In this study, we reported the implementation of the Plackett-Burman (PB) multifactorial design to screen the effects of amino acids on the growth promotion and productivity of a Chinese hamster ovary DG-44 (CHO-DG44) cell line producing bevacizumab. After this screening, the amino acid combinations were optimized by the response surface methodology (RSM) to determine the most effective concentration in feeds. Through this strategy, the... 

Since physiological and pathological processes occur at nano-environments, nanotechnology has considered as an efficient tool for designing of next generation specific biomolecules with enhanced pharmacodynamic and pharmacodynamic properties. In the current investigation, by control of the size and hydrodynamic volume at the nanoscale, for the first time, physicochemical and pharmacokinetic properties of an anti-VEGFA nanobody was remarkably improved by attachment of a Proline-Alanine-Serine (PAS) rich sequence. The results elucidated unexpected impressive effects of PAS sequence on physicochemical properties especially on size, hydrodynamics radius, and even solubility of nanobody. CD... 

The contribution of electrospun nanofibrous membranes (e.NFMs) in the biosensing platforms opens up a new prospect for the invention of faster and more sensitive analytical devices. In this paper, we utilized e.NFM of polyethersulfone (PES) as a solid substrate for the protein immobilization through two different approaches: physical and covalent. Scanning electron microscopy (SEM) and Fourier-transform-infrared (FTIR) tests were performed to study the effect of plasma treatment on protein immobilization efficacy. Moreover, taking advantage of ELISA technique, the influence of different parameters, namely, nanofibers diameter, membrane thickness, plasma treatment time, an incubation time of... 

In this study, the effect of the geometrical parameters of the Lateral Flow Immunoassay (LFI) membrane on the performance of the pregnancy kits is investigated. Consequently, a new geometry for LFI membrane is proposed based on some theoretical, numerical and experimental observations in order to improve the performance of the related kits. A pregnancy kit (for the detection of hCG in urine samples) is developed and the effect of the relocation of the test and the control lines over the membrane was studied based on the variation of the sample fluid velocity. Using Lattice-Boltzmann simulation of the lateral flow within the porous media and the experimental results, the fluid flow within the... 

Vaccines require a period of at least three months for clinical trials, hence a method that can identify elicitation of immune response a few days after the first dose is a necessity. Evolutionary variable selections are modeling approaches for proper manipulation of available data which were used to set up an animal model for classification of time dependent 'HNMR metabolomic profiles and pattern recognition of fluctuations of metabolites in two groups of male rabbits. One group of rabbits was immunized with human red blood cells and the other used as control. Blood was obtained every 48 h from each rabbit for a period of six weeks and the serum monitored for antibodies and metabolites by... 

Endothelial inflammation as a prominent precursor to atherosclerosis elicits a distinct pathological surface expression of particular vascular proteins. To exhibit a site-specific behaviour, micro- and nanoparticles, as carriers of therapeutics or imaging agents, can distinguish and use these proteins as targeted docking sites. Here, a computational patient-specific model capturing the exclusive luminal qualities has been developed to study the transport and adsorption of particles decorated with proper antibodies over an atherosclerotic plaque located in the LAD artery of the patient. Particles, in nano- and micron sizes, have been decorated with Sialyl Lewisx (sLex), P-selectin aptamer... 

Acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also called COVID-19, is one of the most contagious viruses resulting in a progressive pandemic. Since specific antiviral treatments have not been developed yet and its fatal rate is almost high, early and fast detection is critical for controlling the outbreak. In this study, a piezoelectric microcantilever biosensor has been designed for detecting COVID-19 samples directly without requiring preparation steps. The biosensor acts as a transducer and is coated with the related antibody. When the SARS-CoV-2 antigens adsorbed on the microcantilever top surface through their spike proteins, a surface stress due to the mass change would be... 

Pemphigus vulgaris is an autoimmune disease, in which the role of Th17 cytokines needs to be further explored. This study was performed to assess serum levels of three interleukins (IL) required for Th17 differentiation (IL-β, IL-6 and IL-23) and two specific Th17 cytokines (IL- 17 and IL-22) in a group of patients with pemphigus vulgaris, at baseline, 3 weeks and 6 months after of treatment. Correlations between anti-desmogleins and cytokines with disease severity as well as the influence of therapy on the above factors were assessed. Forty-three first-admitted pemphigus vulgaris patients with the active disease entered the study, but only 31 completed the study. Forty-five healthy... 

Electrospray is an old processing technique that has recently been rediscovered. This technology is revolutionizing the biomaterials and nanotechnology fields by opening up new areas of research in fabricating drug delivery nano-devices. Due to their size, nanoparticles have the advantage of reaching less accessible sites in the body by escaping phagocytosis and entering tiny capillaries. Nanoparticles also have the unique property to accumulate at the site of inflammation and therefore, are very suitable for targeted drug delivery. The feasibility of fabricating relatively monodispersed polymeric nanoparticles by electrospray method in a modified electrospray setup is demonstrated in this... 

Background: Extensive full-thickness burns require replacement of both epidermis and dermis. In designing skin replacements, the goal has been to re-create this model and make a product which has both essential components. Methods: In the present study, we developed procedures for establishing confluent, stratified layers of cultured human keratinocytes on the surface of modified collagen-chitosan scaffold that contains fibroblasts. The culture methods for propagation of keratinocytes and fibroblasts isolated from human neonatal foreskin were developed. The growth and proliferation of normal human keratinocytes were evaluated in serum-free (keratinocyte growth medium) and our modified...