BioProcessing Journal

Optimized Protein Yields from Lotus Seed Utilizing CO₂ Pressure Preparation and Ultrasound-Assisted Extraction

The lotus (Nelumbo nucifera) is a fresh-water plant that grows in semitropical climates. It is cultivated extensively in Asian countries, particularly in China. Besides its popularity as an ornamental flower, it is also used as an herbal medicine, mainly in China, Japan, and India, and it displays strong antipyretic, cooling, astringent, and demulcent properties. Lotus seed is widely used as a food in China. Lotus seed is rich in protein, amino acids, unsaturated fatty acids, and has adequate amounts of essential minerals such as potassium, calcium, magnesium, and iron, without heavy metal contamination. As a protein source, the crude protein of lotus seed is higher than parboiled rice (7.70%) and wheat (8.55%). Unlike legumes, lotus seed possesses adequate sulfuric amino acids (methionine + cysteine), comparable to the FAO/WHO and soybean reference patterns. The high content and high quality of protein in lotus seeds emphasizes their value as a vital source of nutrients…

Biologics Production Manufacturing

isoelectric protein precipitation protein extraction protein extraction yield protein precipitation yield salt extraction ultrasound-assisted extraction volatile acid

Cell Culture Bioreactor Optimization: Growing Recombinant HEK293 Cells to Produce a G-Protein-Coupled Receptor (rGPCR) Membrane Protein

Heterologous expression of membrane proteins remains a bottleneck for structural characterization by x-ray crystallography. Such proteins represent approximately 30% of the proteome and are not sufficiently represented in the Protein Data Bank (PDB). G-protein-coupled receptors (GPCRs) are an area of particular interest as it is estimated that one third of current FDA approved drugs act through this class of receptors. We have been studying rhodopsin with an interest in determining the conformational change that leads to signal transduction in this class of receptors. Although there has been some success in expressing select members of the large GPCR family in bacterial systems, the best characterized expression systems have generally been in mammalian tissue culture…

Biologics Production

cell culture bioreactor G-protein-coupled receptors membrane proteins protein expression recombinant proteins stirred-tank bioreactor

Characterization and Qualification of Cell Substrates for Manufacturing Viral Vaccines in the United States

Cell substrates are used in various stages of viral vaccine manufacturing, as in the isolation, selection, and propagation of the virus seed or virus vector stock, as well as for the amplification of the virus to produce the final vaccine product. The various stages of cell substrate use, including cell banking, are shown in a generic manufacturing scheme in Figure 1. Traditionally, viral vaccines have been produced in animal tissues, primary cell cultures, and cell lines that either have a finite life span, such as normal diploid cells, or a theoretically infinite life span, as achieved with continuous or immortalized neoplastic cells. The cell substrates used in viral vaccines currently licensed in the US are listed in Table 1…

Manufacturing Viral Vectors

cell substrates characterization FDA safety testing vaccines viral vaccines

A Summary of the Manufacture, Biochemical Characterization, and Virological Safety Demonstration of the Mouse mAb CB.Hep-1 Used to Produce the Hepatitis B Vaccine

Monoclonal antibodies (mAb) are highly selective molecules, and an unlimited amount of mAbs with equal quality can be produced using mammalian cell cultures and animals. These molecules have remarkable applications in biomedicine, diagnosis and therapy due to the ability to reproduce exactly the same binding properties. The mAbs have been generated against an ostensible set of compounds such as toxins, drugs, blood proteins, cancer cells, viruses, hormones, environmental pollutants, food products, metals and plant materials. In general, mAbs can also be used for creating sensitive tests to detect small amounts of substances, and in therapies, abzymes, and for isolating specific compounds from complex mixtures by immunoaffinity chromatography (IAC)…

Manufacturing

ascites method characterization immunoaffinity chromatography monoclonal antibody vaccines viral clearance viral safety

Profiling Intracellular Monoclonal Antibody Expression for Improved Cell Culture Characterization and Process Optimization

Many laboratories have utilized cell-free systems or prokaryotic systems designed to produce biological molecules with single polypeptide chains, limited folding requirements, and without glycosylation. The yeast systems are used to generate glycoproteins; however, their glycosylation profiles are vastly different from those of mammalian cells. Without significant glycoengineering, the yeast-produced recombinant glycoproteins may be unsuitable as therapeutic molecules. As such, the use of mammalian cells is still the preferred method to produce complex biological molecules…

Biologics Production

antibody expression characterization flow cytometry mammalian cell culture process development specific productivity

Applying Modern Design of Experiments to Mass Transfer Characterization in Bioreactors

Mass transfer is one of the most crucial parameters during scale-up in cell culture, and may cause variations in specific yields. At the cellular level, all the cells require proper supplementation of essential nutrient sources: nitrogen, oxygen, and carbon. The ability of cells to grow, maintain viability, and provide high specific productivity depends on the proper distribution of these nutrients. The latter depends on proper mass transfer to supply oxygen, carbon, and nitrogen through agitation and aeration, along with other micronutrients…

Manufacturing

bioreactor design of experiment mass transfer mass transfer characterization mass transfer coefficient scale-up

Functionally-Improved Recombinant Antibodies: Moving Toward Higher Levels of Expression

Directed evolution has profoundly changed the way antibodies or antigen binding fragments (Fab) are produced today. Lead generation for product candidates is oftentimes based on complex combinatorial libraries containing a large collection of variant antibodies. The selection of candidate molecules by screening procedures like phage display is considered the gold standard. One major advantage of these methods is that initial candidate molecules can be further improved in terms of affinity, specificity, etc. by reiterative in vitro maturation processes using the very methods that have been used for lead generation…

Biologics Production

antibody expression antibody libraries directed evolution gene optimization phage display recombinant antibodies

Peptide Synthesis in Pharmaceutical Manufacturing

Amino acids are the most basic of building blocks in physiology, which is why peptides (oligomers of amino acids) are continuing to grow in prominence among pharmaceutical manufacturers. With inherent abilities to block and/or enhance signal transfers in the human body, peptides, when harnessed as active pharmaceutical ingredients, can treat a host of metabolic diseases, cardiovascular and heart conditions, and neurodegenerative disorders. Peptide-based drug targets are being identified at an increasingly rapid pace, both in terms of recently introduced therapies, and products in the development pipeline…

Manufacturing

peptide synthesis peptide-based therapy purification sequence analysis solid-phase synthesis

Optimization of Baculovirus-Mediated Gene Delivery into Vertebrate Cells

Baculoviruses are large rod-shaped DNA-viruses which specifically infect only arthropods, mostly lepidopteran species. Among the over 600 known species, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the most characterized and widely used. The baculovirus genome is poorly transcribed and does not replicate in non-target cells. Thus, baculoviruses are harmless for vertebrates even though they can be found everywhere and we eat them daily on our vegetables. Baculoviruses have been applied in biotechnology for almost a century now, first as biopesticides (since the 1920s), then for recombinant protein production (since the early 1980s), and most recently for gene delivery into vertebrate cells (since the mid-1990s)…

Baculovirus Expression Technology

baculovirus-mediated gene expression BEVS gene delivery non-target cells

Authentication and Characterization of Animal Cell Lines: The Impact on Research

Propagation and culturing of animal cells is fundamental to biomedical research. Over the past decade, there has been an increased demand for cell lines for usage as both research tools and models by academic and industrial scientists. Cell culture is a critical tool in such areas as cell biology, gene therapy, genomics, transcriptomics and proteomics. The increased demand for cells and cell-based assays has triggered a remarkable boost in cell culture activities, which in turn has lead to a greater incidence of misidentified and contaminated cell cultures…

Research

animal cell line authentication characterization contaminant control microbial contamination morphology

BioProcessing Journal Posts

SEARCH