Measurement of high protein concentrations by optical rotation: a case study for monitoring of monoclonal antibody drug downstream processes

2021 ◽  
Vol 22 ◽  
Author(s):  
Hidenori Inaba ◽  
Kosuke Wakabayashi ◽  
Ikuo Tsujimoto ◽  
Noriko Yoshimoto ◽  
Shuichi Yamamoto
Keyword(s):  
Monoclonal Antibody ◽  
Membrane Filtration ◽  
Optical Rotation ◽  
Protein A ◽  
Small Variation ◽  
Hplc Method ◽  
Unit Operations ◽  
Downstream Processes ◽  
Good Agreement

Background: Recent advancements in cell engineering and bioreactor engineering have enabled high monoclonal antibody (mAb) concentrations in harvested solutions for the downstream process (DSP). Methods: As many unit operations such as capture chromatography, polish chromatography, membrane filtration, virus inactivation, virus filtration, and concentration by ultrafiltration are involved in DSP, it is crucial to monitor the process carefully in order to perform reliable and stable DSP operations. One of the most important signals (process parameter) to be monitored is the protein concentration CP. Although various methods are available, most of them are not suited for measuring high CP. In this paper, we have developed a method for measuring very high CP by optical rotation (OR). Result: Linear correlations were confirmed between OR and CP in the range CP = 0 to 80 g/L for mAbs with high repeatability and small variation coefficients. This method was applied to the monitoring of CP in the opaque (colored) solution during the cell culture. The CP by OR was in good agreement with those by the standard Protein A HPLC method. Conclusion: Monitoring of high CP by OR is expected to be an efficient process analytical tool (PAT) for DSP.

Assay for islet cell antibodies. Protein A--monoclonal antibody method

Diabetes ◽  
1985 ◽  
Vol 34 (3) ◽  
pp. 300-305 ◽  
Author(s):  
S. Srikanta ◽  
A. Rabizadeh ◽  
M. A. Omar ◽  
G. S. Eisenbarth
Keyword(s):  
Monoclonal Antibody ◽  
Islet Cell ◽  
Protein A ◽  
Islet Cell Antibodies ◽  
Antibody Method

Evaluation of the Efficiency of Protein A Affinity Chromatography to Purify a Monoclonal Antibody for Cancer Treatment and its Purity Analysis

2020 ◽  
Vol 7 (2) ◽  
pp. 121-133
Author(s):  
Ayesha Akhtar ◽  
Shivakumar Arumugam ◽  
Shoaib Alam
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Affinity Chromatography ◽  
Protein A ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
High Yield ◽  
Staphylococcal Protein A ◽  
Purification Step ◽  
Purity Analysis

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

scholarly journals Single Chain Variable Fragment Fused to Maltose Binding Protein: A Modular Nanocarrier Platform for the Targeted Delivery of Antitumorals

2021 ◽  
Author(s):  
Francisco J. Reche-Perez ◽  
Simona Plesselova ◽  
Eduardo De los Reyes-Berbel ◽  
Mariano Ortega-Muñoz ◽  
F. Javier Lopez-Jaramillo ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Targeted Delivery ◽  
Specific Binding ◽  
Protein A ◽  
Antibody Fragments ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Binding Properties ◽  
Single Chain Variable Fragments ◽  
Selective Delivery

The use of the specific binding properties of monoclonal antibody fragments such as single-chain variable fragments (ScFv) for the selective delivery of antitumor therapeutics for cancer cells is attractive due...

scholarly journals Comparative environmental impact evaluation using life cycle assessment approach: a case study of integrated membrane-filtration system for the treatment of aerobically-digested palm oil mill effluent

2021 ◽  
Vol 31 (1) ◽  
Author(s):  
Yeit Haan Teow ◽  
Meng Teck Chong ◽  
Kah Chun Ho ◽  
Abdul Wahab Mohammad
Keyword(s):  
Activated Carbon ◽  
Life Cycle ◽  
Palm Oil ◽  
Membrane Filtration ◽  
Electricity Consumption ◽  
Sand Filters ◽  
Unit Operations ◽  
Filtration System ◽  
Pre Treatment ◽  
Aerobically Digested

AbstractAiming to mitigate wastewater pollution arising from the palm oil industry, this university-industry research-and-development project focused on the integration of serial treatment processes, including the use of moving bed biofilm reactor (MBBR), pre-treatment with sand filters and activated carbon filters, and membrane technology for aerobically-digested palm oil mill effluent (POME) treatment. To assess the potential of this sustainable alternative practice in the industry, the developed technology was demonstrated in a pilot-scale facility: four combinations (Combinations I to IV) of unit operations were developed in an integrated membrane-filtration system. Combination I includes a MBBR, pre-treatment unit comprising sand filters and activated carbon filters, ultrafiltration (UF) membrane, and reverse osmosis (RO) membrane, while Combination II excludes MBBR, Combination III excludes UF membrane, and Combination IV excludes both MBBR and UF membrane. Life cycle assessment (LCA) was performed to evaluate potential environmental impacts arising from each combination while achieving the goal of obtaining recycled and reusable water from the aerobically-digested POME treatment. It is reported that electricity consumption is the predominant factor contributing to most of those categories (50–77%) as the emissions of carbon dioxide (CO2), sulfur dioxide (SO2), nitrogen oxides, and volatile mercury during the combustion of fossil fuels. Combination I in the integrated membrane-filtration system with all unit operations incurring high electricity consumption (52 MJ) contributed to the greatest environmental impact. Electricity consumption registers the highest impact towards all life cycle impact categories: 73% on climate change, 80% on terrestrial acidification, 51% on eutrophication, and 43% on human toxicity. Conversely, Combination IV is the most environmentally-friendly process, since it involves only two-unit operations – pre-treatment unit (comprising sand filters and activated carbon filters) and RO membrane unit – and thus incurs the least electricity consumption (41.6 MJ). The LCA offers insights into each combination of the operating process and facilitates both researchers and the industry towards sustainable production.

scholarly journals Model Based Biomass System Design of Feedstock Supply Systems for Bioenergy Production

2013 ◽  
Author(s):  
Kara G. Cafferty ◽  
David J. Muth ◽  
Jacob J. Jacobson ◽  
Kenneth M. Bryden
Keyword(s):  
Supply Chain ◽  
Corn Stover ◽  
Cellulosic Ethanol ◽  
Supply System ◽  
Cellulosic Biomass ◽  
Systems Dynamics ◽  
Climate Conditions ◽  
Unit Operations ◽  
Supply Systems

Engineering feedstock supply systems that deliver affordable, high-quality biomass remains a challenge for the emerging bioenergy industry. Cellulosic biomass is geographically distributed and has diverse physical and chemical properties. Because of this feedstock supply systems that deliver cellulosic biomass resources to biorefineries require integration of a broad set of engineered unit operations. These unit operations include harvest and collection, storage, preprocessing, and transportation processes. Design decisions for each feedstock supply system unit operation impact the engineering design and performance of the other system elements. These interdependencies are further complicated by spatial and temporal variances such as climate conditions and biomass characteristics. This paper develops an integrated model that couples a SQL-based data management engine and systems dynamics models to design and evaluate biomass feedstock supply systems. The integrated model, called the Biomass Logistics Model (BLM), includes a suite of databases that provide 1) engineering performance data for hundreds of equipment systems, 2) spatially explicit labor cost datasets, and 3) local tax and regulation data. The BLM analytic engine is built in the systems dynamics software package Powersim™. The BLM is designed to work with thermochemical and biochemical based biofuel conversion platforms and accommodates a range of cellulosic biomass types (i.e., herbaceous residues, short-rotation woody and herbaceous energy crops, woody residues, algae, etc.). The BLM simulates the flow of biomass through the entire supply chain, tracking changes in feedstock characteristics (i.e., moisture content, dry matter, ash content, and dry bulk density) as influenced by the various operations in the supply chain. By accounting for all of the equipment that comes into contact with biomass from the point of harvest to the throat of the conversion facility and the change in characteristics, the BLM evaluates economic performance of the engineered system, as well as determining energy consumption and green house gas performance of the design. This paper presents a BLM case study delivering corn stover to produce cellulosic ethanol. The case study utilizes the BLM to model the performance of several feedstock supply system designs. The case study also explores the impact of temporal variations in climate conditions to test the sensitivity of the engineering designs. Results from the case study show that under certain conditions corn stover can be delivered to the cellulosic ethanol biorefinery for $35/dry ton.

Structural Analysis of an Offshore Jacket Model Under Seismic Excitation

2008 ◽  
Author(s):  
Helder J. D. Correia ◽  
Anto´nio C. Mendes ◽  
Carlos A. F. S. Oliveira
Keyword(s):  
Point Of View ◽  
Shaking Table ◽  
Seismic Excitation ◽  
Model Structure ◽  
Structural Members ◽  
Jacket Structures ◽  
Reaction Forces ◽  
Nodal Displacements ◽  
Good Agreement

In the present work the action of earthquakes upon offshore jacket structures is analysed by means of ADINA software. Our case-study refers to an existing model structure, previously constructed at the Laboratory of Fluid Mechanics of UBI, which has been analysed from the hydrodynamic point of view — Mendes et al. [1, 2]. The seismic excitation will be imposed at the base of this model structure, with frequencies and amplitudes corresponding to actual earthquake conditions transposed to the model scale of 1:45. The FEM software is utilised to calculate the natural frequencies of the model and to obtain stresses at selected members, as well as their nodal displacements. Our purpose is to quantify maximum stresses occurring in critical structural members and to verify the survivability criterion. The predictions of the numerical model, in terms of the reaction forces at the base and acceleration at the top of the structure, are then correlated with the experimental measurements performed when the model structure is excited in an especially designed shaking table (Correia [3]), revealing a good agreement between both results.

scholarly journals Immunoelectron microscopic localization of an oviductal antigen in hamster zona pellucida by use of a monoclonal antibody.

1988 ◽  
Vol 36 (11) ◽  
pp. 1441-1447 ◽  
Author(s):  
F W Kan ◽  
S St-Jacques ◽  
G Bleau
Keyword(s):  
Monoclonal Antibody ◽  
Zona Pellucida ◽  
Preimplantation Embryo ◽  
Protein A ◽  
Cumulus Cells ◽  
Gold Particles ◽  
Antigenic Sites ◽  
Cumulus Oophorus ◽  
Entire Thickness ◽  
Immunocytochemical Method

The zona pellucida is an extracellular matrix of glycoproteins which surrounds the mammalian oocyte and preimplantation embryo. We have recently developed monoclonal antibodies against oviductal zona pellucida of the golden hamster. We applied the post-embedding immunocytochemical method using a monoclonal antibody (IgGl,k) to determine the precise location of antigenic sites in the cumulus oophorus complex of the superovulated hamster. By applying the high-resolution protein A-gold technique, we demonstrated that the sites of immunoreactivity were exclusively in the zona pellucida encompassing the oocyte. Other structures within the oocyte and neighboring cumulus cells were not labeled by gold particles. Moreover, gold particles were evenly distributed throughout the entire thickness of the zona pellucida, indicating that this extracellular layer is at least in part made up of an antigen recognized by the monoclonal antibody that is uniformly distributed in the zona matrix.

Analysis of Thermal Variation of Length and Refractive Index of Lead Fluoride to Study its Optical Properties

1989 ◽  
Vol 172 ◽  
Author(s):  
T. S. Aurora ◽  
D. O. Pederson ◽  
S. M. Day
Keyword(s):  
Refractive Index ◽  
Room Temperature ◽  
Linear Thermal Expansion ◽  
Small Variation ◽  
Published Data ◽  
Thermal Variation ◽  
Lead Fluoride ◽  
Superionic Phase Transition ◽  
Lorenz Relation ◽  
Good Agreement

AbstractLinear thermal expansion and refractive index variation have been measured in lead fluoride with a laser interferometer as a function of temperature. Data has been analyzed using the Lorentz-Lorenz relation. Molecular polarizability, band gap, variation of refractive index with density, and strain-polarizability parameter have been studied as a function of temperature. They exhibit a small variation with temperature except near the superionic phase transition where the variation appears to be more pronounced. The results are in good agreement with the published data near room temperature.

HPLC METHOD DEVELOPMENT FOR ESTIMATION OF RAMELTEON IN TABLET DOSAGE FORM

INDIAN DRUGS ◽  
2013 ◽  
Vol 50 (06) ◽  
pp. 20-23
Author(s):  
S Sahoo ◽  
◽  
P. K. Panda ◽  
S. K. Mishra
Keyword(s):  
Flow Rate ◽  
Dosage Form ◽  
Method Development ◽  
Hplc Method ◽  
Reverse Phase Hplc ◽  
Ich Guidelines ◽  
Hplc Method Development ◽  
Tablet Dosage ◽  
Good Agreement

A simple, fast, accurate and precise reverse phase HPLC method is developed and described for the determination of ramelteon in tablet dosage form. Chromatography was carried on an ODS column using a mixture of acetonitrile and phosphate buffer pH 7.0 (35:65 V/V) as the mobile phase at a flow rate of 1.0 mL/min with detection at 286 nm. The retention time of the drug was 7.7 min. The procedure was validated for linearity, precision, accuracy, and robustness. The developed method was validated for linearity from 50 to 150% which shows the method is quite linear with a correlation coefficient of 0.999, for precision which includes system precision, method precision, intraday and by another analyst on another day, and accuracy. The %RSD for system precision was observed to be 1.1, whereas the method precision was observed to be 0.2. The % recovery from ‘accuracy’ studies yielded the recovery of 99.7-101.5% which indicates the capability of the method, and finally for robustness that includes studies w.r.t. change in flow rate, the percentage of organic modifier and pH. As per ICH guidelines, method validation results are in good agreement. The proposed method was simple, sensitive, precise and accurate.

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