A comparative study of the bispecific monoclonal antibody,blinatumomab expression in CHO cells and E. coli

Abstract

The “bispecifics” market improved over the past decade due to the development of many technological platforms including bispecific T cell engagers (BiTEs). The approval of blinatumomab, the most advanced bispecific T-cell engager (BiTE) in clinical trials, can be a significant milestone in the development of bispecific antibodies. Both Chinese hamster ovary (CHO) cells and E. coli strain are considered as the most widely used hosts for the large-scale production of therapeutic monoclonal antibodies. Since both of the economic and qualitative aspects of protein production are important in industry, selection of a suitable protein expression system is very critical. The BsAb gene was cloned into the expression vectors FC550A-1, pcDNA3.1 (+), and PET22b and 6?×?His-tagged BsAb then purified on a Ni-NTA chromatography column. Both SDS–PAGE and Western blotting analysis of the purified protein demonstrated that blinatumomab was successfully expressed as a 55?kDa in both expression systems. The antigen-binding properties of blinatumomab were compared in the mammalian system versus Escherichia coli. The results showed that the purified antibody from a mammalian expression system has better binding activity than the one from E. coli host.     

Inheritance of resistance to wheat yellow rust at adult plant stage

In order to investigate on inheritance and gene action for resistance to yellow rust, the resistant line C.B227 was crossed with the susceptible variety Avocet. Parents (P1 and P2) and the resulting F1, F2 and F3 generations were planted in a randomised complete block design with two replications in the field. The plants were inoculated with 70E0A⁺ pathotype of yellow rust in the research station of Gharakhil, Iran, and evaluated for resistance at adult plant stage. Disease severity and infection type of flag leaf were recorded for each single plant and final coefficient of infection was calculated. The results of weighted ANOVA indicated that the difference among the generations was significant (p?<?0.01) for the trait final infection type. Generation mean analysis showed that dominant effect was more important than additive one. The degree of dominance indicated the presence of complete dominance. Additive, dominance and epistasic additive?×?additive [i] effects were important in genetic control of resistance. The results of generation variance analysis were consistent with generation mean analysis.     

Involvement of aberrant regulation of epigenetic mechanisms in the pathogenesis of Parkinson's disease and epigenetic-based therapies

Parkinson's disease (PD) is known as a progressive neurodegenerative disorder associated with the reduction of dopamine-secreting neurons and the formation of Lewy bodies in the substantia nigra and basal ganglia routes. Aging, as well as environmental and genetic factors, are considered as disease risk factors that can make PD as a complex one. Epigenetics means studying heritable changes in gene expression or function, without altering the underlying DNA sequence. Multiple studies have shown the association of epigenetic variations with onset or progression of various types of diseases. DNA methylation, posttranslational modifications of histones and presence of microRNA (miRNA) are among epigenetic processes involved in regulating pathways related to the development of PD. Unlike genetic mutations, most epigenetic variations may be reversible or preventable. Therefore, the return of aberrant epigenetic events in different cells is a growing therapeutic approach to treatment or prevention. Currently, there are several methods for treating PD patients, the most important of which are drug therapies. However, detection of genes and epigenetic mechanisms involved in the disease can develop appropriate diagnosis and treatment of the disease before the onset of disabilities and resulting complications. The main purpose of this study was to review the most important epigenetic molecular mechanisms, epigenetic variations in PD, and epigenetic-based therapies.     

Systematic Analysis of Protein–Protein and Gene–Environment Interactions to Decipher the Cognitive Mechanisms of Autism Spectrum Disorder

Cellular and Molecular Neurobiology - Autism spectrum disorder (ASD), a heterogeneous neurodevelopmental disorder resulting from both genetic and environmental risk factors, is manifested by...     

Genetics of Familial Combined Hyperlipidemia (FCHL) Disorder: An Update

Biochemical Genetics - Familial combined hyperlipidemia (FCHL) is one of the most common familial lipoprotein disorders of the lipoproteins, with a prevalence of 0.5% to 2% in different...     

Distance learning techniques for ontology similarity measuring and ontology mapping

Recent years, a large amount of ontology learning algorithms have been applied in different disciplines and engineering. The ontology model is presented as a graph and the key of ontology algorithms is similarity measuring between concepts. In the learning frameworks, the information of each ontology vertex is expressed as a vector, thus the similarity measuring can be determined via the distance of the corresponding vector. In this paper, we study how to get an optimal distance function in the ontology setting. The tricks we presented are divided into two parts: first, the ontology distance learning technology in the setting that the ontology data have no labels; then, the distance learning approaches in the setting that the given ontology data are carrying real numbers as their labels. The result data of the four simulation experiments reveal that our new ontology trick has high efficiency and accuracy in ontology similarity measure and ontology mapping in special engineering applications.     

Nigrostriatal reduction of aromatic L-amino acid decarboxylase activity in MPTP-treated squirrel monkeys: in vivo and in vitro investigations

Aromatic L-amino acid decarboxylase (AAAD) activity was examined in vivo with positron emission tomography (PET) using 6-[18F]fluoro-L-DOPA (FDOPA) in squirrel monkeys lesioned with graded doses of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). In vitro biochemical determinations of AAAD activity in caudate, putamen, substantia nigra, and nucleus accumbens were performed in the same animals to establish a direct comparison of in vivo and in vitro measurements. In vivo and in vitro AAAD activities in caudate/ putamen were substantially reduced in animals treated with the highest dose of MPTP (2.0 mg/kg). The percent change in the striatal FDOPA uptake (K(i)) and decarboxylation rate constant (k3) values resulting from MPTP treatment showed highly significant correlations with in vitro-determined AAAD activities. However, decarboxylase rates within individual animals presented as approximately 10-fold difference between in vivo and in vitro values. Lower in vivo k3 measurements may be attributed to several possibilities, including transport restrictions limiting substrate availability to AAAD within the neuron. In addition, reductions in AAAD activity in the substantia nigra did not parallel reductions in AAAD activity within the striatum, supporting the notion of a nonlinear relationship between nigrostriatal cell degeneration and terminal losses. This work further explores the role of AAAD in Parkinson's disease, a more important factor than previously thought.     

Biology of Apanteles myeloenta (Hymenoptera: Braconidae), a larval parasitoid of carob moth Ectomyelais ceratoniae (Lepidoptera: Pyralidae)

The carob moth, Ectomyelois ceratoniae (Lepidoptera: Pyralidae), is the most important pest of pomegranate orchards (in terms of economic damage) within Iran, and hence, several control procedures, including biological methods of control, have been attempted as a means of controlling populations of this insect. This research was carried out in order to study the biology of Apanteles myeloenta (Hymenoptera: Braconidae), a larval parasitoid of the carob moth. Laboratory studies were conducted to determine larval developmental time, adult longevity, sex ratio, parasite progeny production, and host stage preference of A. myeloenta. At 25 ± 1 °C, immature developmental time (egg to pupa; mean ± SE) was 28.33 ± 0.85 days and 27.46 ± 0.37 days for male and females, respectively. Adult females survived on average 17.5 ± 0.14, 11.7 ± 0.22, 3.4 ± 0.18, and 2.8 ± 0.12 days at 25 C when provided with honey and water, honey only, water only or no food source, respectively. The sex ratio (females to males) of A. myeloenta was 1:3.5 from hosts parasitized in the first instar, 1:3 for second instars and 1:2 for third instar carob moth larvae. Female A. myeloenta typically preferred to parasitize second instar over third or first instar. The oviposition activity peaked on the 7th and 8th days following emergence, when provided with honey, and 10% sucrose solution, respectively.     

The Quantitative Imaging Network: NCI's Historical Perspective and Planned Goals

The purpose of this editorial is to provide a brief history of National Institutes of Health National Cancer Institute (NCI) workshops as related to quantitative imaging within the oncology setting. The editorial will then focus on the recently supported NCI initiatives, including the Quantitative Imaging Network (QIN) initiative and its organizational structure, including planned research goals and deliverables. The publications in this issue of Translational Oncology come from many of the current members of this QIN research network.     

Infra-specific genetic and morphological diversity in Linum album (Linaceae)

The genus Linum L. (Lineacea) has over 15 species, subspecies or ecotypes in Iran. These species show extensive geographical distribution and form many local populations throughout the country. Linum album is herbaceous medicinal plant containing important lignans such as podophyllotoxin (PTOX) and 6-methoxy podophyllotoxin (MPTOX), which have antiviral and anticancer properties. Studying the genetic and morphological diversity of different geographical populations produces detailed knowledge about population divergence and identification of the infra-species taxa if at all they are present. Moreover, the populations that differ in their genetic content and structure may also differ in their chemical and medicinal properties. The present study considers morphological and genetic diversity analyses of 20 L. album geographical populations by using nuclear ISSR markers, genome size, and cytogenetic characteristics. These populations differed significantly in many of their quantitative morphological characters and in some of their qualitative features. They also differed significantly in their molecular characteristics and genome size. Details of morphological and molecular variations are reported and discussed.