scholarly journals Affinity Chromatography: An Enabling Technology for Large Scale Bioprocessing - How does it get to the Developing World

2021 ◽  
Vol 2 (7) ◽  
pp. 01-02
Author(s):  
Frank Riske ◽  
Brendan Riske
Keyword(s):  
Affinity Chromatography ◽  
Large Scale ◽  
Developing World ◽  
Process Development ◽  
Enzyme Inhibitor ◽  
Specific Interaction ◽  
Protein A ◽  
Biological Interactions ◽  
Antibody Capture ◽  
Developed World

Affinity chromatography was initially used to describe chromatographic biological interactions such as lectin-glycoprotein, antibody -antigen and enzyme-inhibitor. This definition has expanded to include the specific interaction between a target and a ligand. The use of affinity chromatography has reached a zenith with the explosion of Mab therapeutics and the use of Protein-A chromatography for antibody capture. Now, affinity chromatography has moved to non Mab proteins. This can result in the same economic advantages as Mab, by enabling the standardization of process development and manufacturing processes in flexible multiproduct production sites. The output is improved product throughput, higher target recoveries, and potentially less expensive drugs. These advantages are available to the developed world but how do we make this technology available to the developing world?

LARGE SCALE PREPARATION OF VON WILLEBRAND FACTOR BY AFFINITY CHROMATOGRAPHY

1987 ◽  
Author(s):  
J Newman ◽  
D Farb
Keyword(s):  
Factor Viii ◽  
Affinity Chromatography ◽  
Von Willebrand Factor ◽  
Large Scale ◽  
Specific Activity ◽  
Protein A ◽  
Sephadex Column ◽  
Scale Preparation ◽  
Von Willebrand ◽  
Willebrand Factor

Treatment of bleeding in von Willebrand's disease usually consists of the infusion of cryoprecipitate or plasma. DDAVP is effective in some patients. Commercial concentrates for the treatment of Hemophilia A are ineffective as a source of von Willebrand Factor (vWF) replacement in von Willibrand's disease presumedly because of the absence of higher molecular weight forms of vWF protein. A vWF concentrate obtained during the course of preparation of an affinity purified Factor VIII may provide an alternative therapeutic agent without impacting the available Factor VIII supplies. The process used for the preparation of a highly purified Factor VIII concentrate (MonoclateTM, Armour Pharmaceutical Co.) from cryoprecipitate includes an affinity chromatography step which separates vWF/Factor VIII complex from other proteins in cryoprecipitate using an anti-vWF monoclonal antibody (C. A. Fulcher & T. S. Zimmerman, 1982). Factor VIII is then dissociated from the vWF remaining on the column and is eluted immediately by 3M sodium thiocyanante (NaSCN) as a step in the regeneration of the column. Unless the NaSCN is rapidly removed from the elutriant, the vWF activity as measured by platelet agglutination is destroyed. We have taken the NaSCN eluate and processed it immediately over an in-line G-10 or G-25 Sephadex column which removes the NaSCN while the vWF is eluted with a buffered isotonic solution. Alternatively, the vWF has been precipitated from the NaSCN by ammonium sulfate or polyethylene glycol. VWF prepared by any of these methods retains platelet agglutinating activity and has a distribution of vWF multimers similar to those of vWF in normal plasma.The potency of vWF prepared from cryoprecipitate by this process is 20 units/ml and the specific activity is 20 units/mg. In several fractionations of kilogram amounts of cryoprecipitate, vWF was isolated and subjected to lyophilization and heating at 68° for 30 hours without loss of bioactivity. Drug development in progress suggest that the combined effect of affinity chromatography and exposure to NaSCN may negate the need for a heating procedure to reduce the risk of viral transmittance.

scholarly journals Applications of Technology for Livestock for Small Holders to Meet Global Food Production

Ceiba ◽  
2016 ◽  
Vol 54 (1) ◽  
pp. 3-13
Author(s):  
Max F. Rothschild ◽  
Graham Plastow
Keyword(s):  
Food Production ◽  
Large Scale ◽  
Developing World ◽  
Green Revolution ◽  
Reproductive Technologies ◽  
Livestock Production ◽  
Animal Products ◽  
Balanced Diet ◽  
Small Holders ◽  
Developed World

While the Green Revolution greatly changed production of crops worldwide and helped feed over a billion people, improved production of livestock has been more limited across the globe. Modern advances in livestock production have in general only benefitted two groups: large scale livestock producers and consumers in the developed world. In some parts of the world many of the animal production practices have not changed for the last 1000 years and in other regions small holders have benefited only marginally by the scientific advances that now are an integral part of large scale commercial production. However, increased food insecurity and a worldwide food production crisis loom in the future as the most significant scientific challenge facing us in the next 30 years. Expectations are that human population growth will soon go from 7.3 billion to 9.6 billion by 2050, and food production must increase rapidly to meet the demand. These increases must come despite evidence of climate change and limited land and water resources. Whilst there is a perception of over consumption of animal products in the developed world, there are still significant numbers that are undernourished. Animal protein plays a very important role in achieving a balanced diet in the developing world, especially for the adequate nutrition of children. Furthermore, it is expected that significant increased demand for animal source foods will occur especially in the poorer countries of the developing world where most livestock are produced by small holders, but also in countries like China and Brazil which are transforming quickly. Many researchers have touted that modern feeding systems, and the use of advanced reproductive technologies and advanced genetics and genomics will provide solutions to increasing food in the developing world. These opportunities certainly exist, but direction and focus of research, funding issues, human capacity training and training of small holders will all be required for increasing livestock production to satisfy these demands. These activities will need to be embedded within sustainable programs that address implementation from the outset, and benefiting small holder production will be crucial to meeting this challenge.

The Structure and Properties of MoSi2 Thin Film in Mos Process

1980 ◽  
Vol 38 ◽  
pp. 326-327
Author(s):  
C.K. Wu ◽  
P. Chang ◽  
N. Godinho
Keyword(s):  
Thin Film ◽  
Integrated Circuits ◽  
High Performance ◽  
Large Scale ◽  
Process Development ◽  
Structure And Properties ◽  
Metal Silicides ◽  
High Oxidation ◽  
Important Approach ◽  
High Oxidation Resistance

Recently, the use of refractory metal silicides as low resistivity, high temperature and high oxidation resistance gate materials in large scale integrated circuits (LSI) has become an important approach in advanced MOS process development (1). This research is a systematic study on the structure and properties of molybdenum silicide thin film and its applicability to high performance LSI fabrication.

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 Functionalization of Cellulose-Based Hydrogels with Bi-Functional Fusion Proteins Containing Carbohydrate-Binding Modules

Materials ◽  
2021 ◽  
Vol 14 (12) ◽  
pp. 3175
Author(s):  
Mariana Barbosa ◽  
Hélvio Simões ◽  
Duarte Miguel F. Prazeres
Keyword(s):  
Fusion Proteins ◽  
Fluorescent Protein ◽  
Protein A ◽  
Chemical Properties ◽  
Main Idea ◽  
Bioactive Molecules ◽  
Scaffolding Protein ◽  
Carbohydrate Binding ◽  
Biological Interactions ◽  
Carbohydrate Binding Modules

Materials with novel and enhanced functionalities can be obtained by modifying cellulose with a range of biomolecules. This functionalization can deliver tailored cellulose-based materials with enhanced physical and chemical properties and control of biological interactions that match specific applications. One of the foundations for the success of such biomaterials is to efficiently control the capacity to combine relevant biomolecules into cellulose materials in such a way that the desired functionality is attained. In this context, our main goal was to develop bi-functional biomolecular constructs for the precise modification of cellulose hydrogels with bioactive molecules of interest. The main idea was to use biomolecular engineering techniques to generate and purify different recombinant fusions of carbohydrate binding modules (CBMs) with significant biological entities. Specifically, CBM-based fusions were designed to enable the bridging of proteins or oligonucleotides with cellulose hydrogels. The work focused on constructs that combine a family 3 CBM derived from the cellulosomal-scaffolding protein A from Clostridium thermocellum (CBM3) with the following: (i) an N-terminal green fluorescent protein (GFP) domain (GFP-CBM3); (ii) a double Z domain that recognizes IgG antibodies; and (iii) a C-terminal cysteine (CBM3C). The ability of the CBM fusions to bind and/or anchor their counterparts onto the surface of cellulose hydrogels was evaluated with pull-down assays. Capture of GFP-CBM3 by cellulose was first demonstrated qualitatively by fluorescence microscopy. The binding of the fusion proteins, the capture of antibodies (by ZZ-CBM3), and the grafting of an oligonucleotide (to CBM3C) were successfully demonstrated. The bioactive cellulose platform described here enables the precise anchoring of different biomolecules onto cellulose hydrogels and could contribute significatively to the development of advanced medical diagnostic sensors or specialized biomaterials, among others.

scholarly journals Peptide Blocking CTLA-4 and B7-1 Interaction

Molecules ◽  
2021 ◽  
Vol 26 (2) ◽  
pp. 253
Author(s):  
Stepan V. Podlesnykh ◽  
Kristina E. Abramova ◽  
Anastasia Gordeeva ◽  
Andrei I. Khlebnikov ◽  
Andrei I. Chapoval
Keyword(s):  
Cancer Immunotherapy ◽  
Functional Activity ◽  
Synthetic Peptide ◽  
Checkpoint Inhibitors ◽  
Specific Interaction ◽  
Protein A ◽  
Tumor Rejection ◽  
Immune Checkpoints ◽  
3D Computer Model ◽  
Molecular Compounds

Discovery of the B7 family immune checkpoints such as CTLA-4 (CD152), PD-1 (CD279), as well as their ligands B7-1 (CD80), B7-2 (CD86), B7-H1 (PD-L1, CD274), and B7-DC (PD-L2, CD273), has opened new possibilities for cancer immunotherapy using monoclonal antibodies (mAb). The blockade of inhibitory receptors (CTLA-4 and PD-1) with specific mAb results in the activation of cancer patients’ T lymphocytes and tumor rejection. However, the use of mAb in clinics has several limitations including side effects and cost of treatment. The development of new low-molecular compounds that block immune checkpoints’ functional activity can help to overcome some of these limitations. In this paper, we describe a synthetic peptide (p344) containing 14 amino acids that specifically interact with CTLA-4 protein. A 3D computer model suggests that this peptide binds to the 99MYPPPY104 loop of CTLA-4 protein and potentially blocks the contact of CTLA-4 receptor with B7-1 ligand. Experimental data confirm the peptide-specific interaction with CTLA-4 and its ability to partially block CTLA-4/B7-1 binding. The identified synthetic peptide can be used for the development of novel immune checkpoint inhibitors that can block CTLA-4 functional activity for cancer immunotherapy.

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