scholarly journals Tuning Relative Polypeptide Expression to Optimize Assembly, Yield and Downstream Processing of Bispecific Antibodies

Antibodies ◽  
2018 ◽  
Vol 7 (3) ◽  
pp. 29 ◽  
Author(s):  
Giovanni Magistrelli ◽  
Guillemette Pontini ◽  
Yves Poitevin ◽  
Pauline Malinge ◽  
Jérémie Bourguignon ◽  
...  
Keyword(s):  
Large Scale ◽  
Downstream Processing ◽  
Bispecific Antibodies ◽  
Parallel Optimization ◽  
Optimal Codons ◽  
Antibody Titers ◽  
Constant Domains ◽  
Polypeptide Chains ◽  
Optimal Assembly ◽  
By Products

Bispecific antibodies (bsAbs) are often composed of several polypeptide chains that have to be expressed adequately to enable optimal assembly and yield of the bsAb. κλ bodies are a bispecific format with a native IgG structure, composed of two different light chains that pair with a common heavy chain. Introduction of non-optimal codons into the sequence of a particular polypeptide is an effective strategy for down modulating its expression. Here we applied this strategy but restricted the modification of the codon content to the constant domain of one light chain. This approach facilitates parallel optimization of several bsAbs by using the same modified constant domains. Partial sequence de-optimization reduced expression of the targeted polypeptide. Stable cell pools could be isolated displaying increased bispecific antibody titers as well as changes in the abundance of undesired by-products that require elimination during downstream processing. Thus, modulating the relative expression of polypeptides can have a significant impact on bsAb titer and product related impurities; which are important factors for large scale manufacturing for clinical supply.

Recent Developments in the Downstream Processing of Phycobiliproteins from Algae: A Review

2021 ◽  
Vol 06 ◽  
Author(s):  
Ayekpam Chandralekha Devi ◽  
G. K. Hamsavi ◽  
Simran Sahota ◽  
Rochak Mittal ◽  
Hrishikesh A. Tavanandi ◽  
...  
Keyword(s):  
Cell Wall ◽  
Large Scale ◽  
Downstream Processing ◽  
Review Article ◽  
Current Status ◽  
Wall Structure ◽  
Scale Production ◽  
Cell Wall Disruption ◽  
Recent Developments ◽  
Macro Algae

Abstract: Algae (both micro and macro) have gained huge attention in the recent past for their high commercial value products. They are the source of various biomolecules of commercial applications ranging from nutraceuticals to fuels. Phycobiliproteins are one such high value low volume compounds which are mainly obtained from micro and macro algae. In order to tap the bioresource, a significant amount of work has been carried out for large scale production of algal biomass. However, work on downstream processing aspects of phycobiliproteins (PBPs) from algae is scarce, especially in case of macroalgae. There are several difficulties in cell wall disruption of both micro and macro algae because of their cell wall structure and compositions. At the same time, there are several challenges in the purification of phycobiliproteins. The current review article focuses on the recent developments in downstream processing of phycobiliproteins (mainly phycocyanins and phycoerythrins) from micro and macroalgae. The current status, the recent advancements and potential technologies (that are under development) are summarised in this review article besides providing future directions for the present research area.

scholarly journals Bispecific antibodies with Fab-arms featuring exchanged antigen-binding constant domains

2021 ◽  
Vol 26 ◽  
pp. 100959
Author(s):  
Filippo Benedetti ◽  
Florian Stracke ◽  
Gerhard Stadlmayr ◽  
Katharina Stadlbauer ◽  
Florian Rüker ◽  
...  
Keyword(s):  
Binding Constant ◽  
Bispecific Antibodies ◽  
Antigen Binding ◽  
Constant Domains

The Croonian Lecture, 1970 The structural basis of muscular contraction

1971 ◽  
Vol 178 (1051) ◽  
pp. 131-149 ◽  
Keyword(s):  
Large Scale ◽  
Muscular Contraction ◽  
Important Change ◽  
Structural Basis ◽  
X Ray Diffraction ◽  
New Era ◽  
X Ray ◽  
Biology I ◽  
The Times ◽  
Polypeptide Chains

A previous occasion on which the Croonian lecture was directly concerned with the mechanism of muscular contraction was in 1945, when it was delivered by Professor W. T. Astbury. On that occasion he commented that it was a sign of the times that a physicist should be asked to give the Croonian lecture, and went on to say, and I quote: ‘We are at the dawn of a new era, the era of “molecular biology”, as I like to call it, and there is an urgency about the need for more intensive application of physics and chemistry, and specially structural analysis, to biological problems.’ These were very prophetic words, and, as a physicist just entering biology, I was much interested to read them, and to learn about his experiments. The basic experimental finding which Astbury reported (1947) was that there did not seem to be any important change in the wide angle X-ray diagram from muscle upon contraction; hence it followed that muscles did not contract by any process which simply involved the large-scale disorientation of originally well-ordered polypeptide chains, nor by an alteration in chain configuration in the well-ordered parts of the structure. Astbury suggested instead that there might be ‘specifically active foci’ which one could perhaps paraphrase as ‘larger structural units’ (i.e. larger than individual polypeptide chains) concerned in contraction, which might be studied in the electron microscope or by low angle X-ray diffraction.

scholarly journals HEAVY CLASTIC MINERALS AS AN INDICATOR OF GEODYNAMIC SETTINGS OF ACCUMULATION AND PROVENANCE OF CRETACEOUS SEDIMENTS OF THE WEST SAKHALIN TERRANE

2021 ◽  
pp. 68-86
Author(s):  
A.I. Malinovsky ◽  
Keyword(s):  
Large Scale ◽  
Volcanic Rocks ◽  
Source Area ◽  
Heavy Minerals ◽  
Clastic Material ◽  
The West ◽  
Geodynamic Settings ◽  
Eurasian Continent ◽  
Ocean Boundary ◽  
By Products

The article discusses the results of studying heavy clastic minerals from the Cretaceous sandy rocks of the West Sakhalin Terrane, and also presents their paleogeodynamic interpretation. It is shown that in terms of mineralogical and petrographic parameters, the terrane sandstones correspond to typical graywackes and are petrogenic rocks formed mainly by destruction of igneous rocks of the source areas. The sediments were found to contain both sialic, granite-metamorphic association minerals, and femic, formed by products of the destruction of basic and ultrabasic volcanic rocks. The interpretation of the entire set of data on the content, distribution and microchemical composition of heavy minerals was carried out by comparing them with minerals from older rocks and modern sediments accumulated in known geodynamic settings. The results obtained indicate that during the Cretaceous, sedimentation occurred along the continent-ocean boundary in a basin associated with large-scale left-lateral transform movements of the Izanagi Plate relative to the Eurasian continent. The source area that supplied clastic material to that basin combined a sialic landmass composed of granite-metamorphic and sedimentary rocks, a mature deeply dissected ensialic island arc, and fragments of accretion prisms, in the structure of which involved ophiolites.

scholarly journals FrustratometeR: an R-package to compute Local frustration in protein structures, point mutants and MD simulations

2020 ◽  
Author(s):  
Atilio O. Rausch ◽  
Maria I. Freiberger ◽  
Cesar O. Leonetti ◽  
Diego M. Luna ◽  
Leandro G. Radusky ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Large Scale ◽  
Protein Structures ◽  
Md Simulations ◽  
R Package ◽  
Protein Protein Interactions ◽  
Large Scale Analysis ◽  
Functional Aspects ◽  
Catalytic Sites ◽  
Polypeptide Chains

Once folded natural protein molecules have few energetic conflicts within their polypeptide chains. Many protein structures do however contain regions where energetic conflicts remain after folding, i.e. they have highly frustrated regions. These regions, kept in place over evolutionary and physiological timescales, are related to several functional aspects of natural proteins such as protein-protein interactions, small ligand recognition, catalytic sites and allostery. Here we present FrustratometeR, an R package that easily computes local energetic frustration on a personal computer or a cluster. This package facilitates large scale analysis of local frustration, point mutants and MD trajectories, allowing straightforward integration of local frustration analysis in to pipelines for protein structural analysis.Availability and implementation: https://github.com/proteinphysiologylab/frustratometeR

scholarly journals Expression and manufacturing of protein therapeutics in spirulina

2021 ◽  
Author(s):  
Benjamin Jester ◽  
Hui Zhao ◽  
Mesfin Gewe ◽  
Thomas Adame ◽  
Lisa Perruzza ◽  
...  
Keyword(s):  
Large Scale ◽  
Genetic Manipulation ◽  
Low Cost ◽  
Downstream Processing ◽  
Arthrospira Platensis ◽  
Protein Therapeutics ◽  
High Productivity ◽  
Medical Needs ◽  
Manufacturing Platform ◽  
Safe Food

ABSTRACTArthrospira platensis (commonly known as spirulina) is a photosynthetic cyanobacterium1. It is a highly nutritious food that has been consumed for decades in the US, and even longer by indigenous cultures2. Its widespread use as a safe food source and proven scalability have driven frequent attempts to convert it into a biomanufacturing platform. But these were repeatedly frustrated by spirulina’s genetic intractability. We report here efficient and versatile genetic engineering methodology for spirulina that allows stable expression of bioactive protein therapeutics at high levels. We further describe large-scale, indoor cultivation and downstream processing methods appropriate for the manufacturing of biopharmaceuticals in spirulina. The potential of the platform is illustrated by pre-clinical development and human testing of an orally delivered antibody therapeutic against campylobacter, a major cause of infant mortality in the developing world and a growing antibiotic resistance threat3,4. This integrated development and manufacturing platform blends the safety of food-based biotechnology with the ease of genetic manipulation, rapid growth rates and high productivity characteristic of microbial platforms. These features combine for exceptionally low-cost production of biopharmaceuticals to address medical needs that are unfeasible with current biotechnology platforms.

scholarly journals Conversion of Fructose to HMF in a Continuous Fixed Bed Reactor with Outstanding Selectivity

Molecules ◽  
2018 ◽  
Vol 23 (7) ◽  
pp. 1802 ◽  
Author(s):  
Eric Weingart ◽  
Sarah Tschirner ◽  
Linda Teevs ◽  
Ulf Prüße
Keyword(s):  
Large Scale ◽  
Continuous Process ◽  
Fixed Bed ◽  
Fixed Bed Reactor ◽  
Water Addition ◽  
Batch Systems ◽  
Formation Conditions ◽  
High Yields ◽  
Optimized Conditions ◽  
By Products

5-Hydroxymethylfurfural (HMF) is a very promising component for bio-based plastics. Efficient synthesis of HMF from biomass is still challenging because of fast degradation of HMF to by-products under formation conditions. Therefore, different studies, conducted mainly in monophasic and biphasic batch systems with and without water addition have been published and are still under investigation. However, to produce HMF at a large scale, a continuous process is preferable. Until now, only a few studies have been published in this context. In this work, it is shown that fluorous alcohol hexafluoroisopropanol (HFIP) can act as superior reaction solvent for HMF synthesis from fructose in a fixed bed reactor. Very high yields of 76% HMF can be achieved in this system under optimized conditions, whilst the catalyst is very stable over several days. Such high yields are only described elsewhere with high boiling reaction solvents like dimethylsulfoxide (DMSO), whereas HFIP with a boiling point of 58 °C is very easy to separate from HMF.

scholarly journals Evidence of exposure to SARS-CoV-2 in cats and dogs from households in Italy

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
E. I. Patterson ◽  
G. Elia ◽  
A. Grassi ◽  
A. Giordano ◽  
C. Desario ◽  
...  
Keyword(s):  
Risk Factors ◽  
Large Scale ◽  
Companion Animals ◽  
Neutralizing Antibody ◽  
Northern Italy ◽  
Human Infection ◽  
Pet Ownership ◽  
Natural Conditions ◽  
Factors Associated ◽  
Antibody Titers

AbstractSARS-CoV-2 emerged from animals and is now easily transmitted between people. Sporadic detection of natural cases in animals alongside successful experimental infections of pets, such as cats, ferrets and dogs, raises questions about the susceptibility of animals under natural conditions of pet ownership. Here, we report a large-scale study to assess SARS-CoV-2 infection in 919 companion animals living in northern Italy, sampled at a time of frequent human infection. No animals tested PCR positive. However, 3.3% of dogs and 5.8% of cats had measurable SARS-CoV-2 neutralizing antibody titers, with dogs from COVID-19 positive households being significantly more likely to test positive than those from COVID-19 negative households. Understanding risk factors associated with this and their potential to infect other species requires urgent investigation.

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