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A New Cell-free Protein Expression System with three-fold higher protein yield in batch and continuous mode than existing systems

Researchers at the University of California, Davis have developed a method for preparing a bacterial cell lysate that results in higher protein expression than existing cell-free systems. The new whole-cell lysate system comes with additional advantages, including the ability to synthesize protein from linear DNA, directly amenable to continuous or flow-based reaction, and compatibility with existing manufacturing workflow.

Methods To Suppress Viral Infection Of Mammalian Cells

To meet the ever-growing demand for effective, safe, and affordable protein therapeutics, decades of intense efforts have aimed to maximize the quantity and quality of recombinant proteins produced in Chinese hamster ovary (CHO) cells. CHO cells are extensively used to produce biopharmaceuticals and one advantage is their reduced susceptibility to many human virus families. However, there have been a few episodes of animal viral contamination of biopharmaceutical production runs, mostly from trace levels of viruses in raw materials. These infections more often caused by RNA viruses have led to expensive decontamination efforts and threatened the supply of critical drugs. Viral contamination in biopharmaceutical manufacturing can lead to shortages in the supply of critical therapeutics. Therefore there is a need to understand the mechanisms by which CHO cells are infected and how the cells can be universally engineered to enhance their viral resistance.

Chemoselective Side-Chain Modifications Of Methionine-Containing Elastin-Like Polypeptides

UCLA researchers in the Department of Bioengineering and Department of Chemistry & Biochemistry have developed a novel method for the introduction of various functional groups onto recombinant elastin-like polypeptides (ELPs), creating new compositions of ELPs that may be used for medical therapeutic or diagnostic applications.

Protein Translation Machinery One Shot (TraMOS) Tool

Researchers at the University of California, Davis have developed a microbial culture capable of translating mRNA molecules into a polypeptide with a single reaction mixture.

A Highly Error-Prone Orthogonal Replication System For Targeted Continuous Evolution In Vivo

Inventors at UC Irvine have engineered an orthogonal DNA replication system capable of rapid, accelerated continuous evolution. This system enables the directed evolution of specific biomolecules towards user-defined functions and is applicable to problems of protein, enzyme, and metabolic pathway engineering.

Living Bioreactor for Stoichiometric Protein Production

Living bioreactors are powerful systems for producing a variety of valuable compounds. The versatility of such bioreactors is one of the more useful aspects of the system. Large quantities of compounds or cellular components can be produced efficiently, with minimal cost. Alternately, these systems can be used to produce pathway components that are necessary in the production of secondary products. A common problem with such systems is that they are limited by non-uniform production of pathway components, or require an isolation process to ensure the components are in the appropriate quantity and sequence in the process. Inventors at Texas A&M and UC San Francisco have developed a novel technique to address these issues. The technology effectively results in a stoichiometric production of protein components that are produced in an array, ready for secondary production.

Preparation Of Functional Homocysteine Residues In Polypeptides And Peptides

UCLA researchers in the Department of Bioengineering and Department of Chemistry & Biochemistry have developed a novel method for efficient, chemoselective transformation of methionines in peptides and polypeptides into stable, functional homocysteine derivatives. This method provides a means of creation of new functional biopolymers, site-specific peptide tagging, and synthesis of biomimetic and structural analogs of peptides.

In Situ Lipid Synthesis for Protein Reconstitution (INSYRT)

While current methods for membrane protein functional reconstitution in biomimetic membranes approaches are powerful and have uncovered fundamental properties of protein function, they are methodologically cumbersome, requiring chromatography steps to remove detergents. Moreover, structural features normally found in cell membranes such as curvature and polarity are mostly absent. In this regard, an efficient reconstitution methodology that better mimics the native chemical environment of a whole-cell embedded protein would be highly useful.

Use of Mutant Kv7.2 Channels for Anti-Epileptic and Pain Therapies

During seizures or pain-induced inflammation, excess chemical mediators suppress potassium channels mediating neuronal activity and thereby inactivate new generation anti-epileptic drugs and painkillers acting on those channels. The invention describes a gene therapy using a genetically-engineered potassium channel that reduces adverse effects by silencing neuronal hyperactivity while maintaining normal neuronal activity in the presence of chemical mediators to treat epilepsy and pain.

An Antibody to Phospho T3 of Human Huntingtin

Huntington’s disease (HD) is a neurodegenerative genetic disorder caused by abnormal function of mutated Huntingtin protein. The invention uncovers an antibody to a new post-translational modification site that affects human Huntingtin aggregation and pathogenesis of HD.

DNA Amplification by Electric Field Cycling (efc-PCR)

Polymerase Chain Reaction (PCR) is a popular technique for amplifying and quantifying minute quantities of DNA. Technologies based on PCR are used for a wide range of applications, including forensics, disease detection, and laboratory tools. Researchers at UCI have developed a device that can implement a novel method for PCR based on voltage cycling as opposed to temperature cycling (the current method for PCR). This allows the device to be much more portable and compact than those currently available.

Improved Cell-Free Protein Synthesis For Protein Microarray

Researchers at UC Irvine have developed a cell-free (CF) protein synthesis system to solubilize and synthesize highly hydrophobic membrane proteins that would typically aggregate using current CF synthesis systems. With such high amounts of synthesized proteins, researchers intend to build protein microarrays for diagnostic purposes.

Therapeutic strategies for Huntington’s Disease using stop codon suppression

In Huntington’s Disease (HD), aberrant splicing of the huntingtin protein can produce a highly toxic peptide that accumulates in the brain. The invention describes methods to minimize the toxicity of spliced proteins.

Mammalian Cell Culture Optimization

Biotherapeutic proteins manufactured in cell culture systems have transformed modern medicine. Selling many tens of billions per year, new biotherapeutics such as monoclonal antibodies have delivered dramatic clinical results, while posing significant manufacturing problems.: During the cell culture manufacturing process, toxic bioproducts such as lactate and ammonia have posed considerable challenges in bioprocessing, since they limit cell growth and impact critical quality attributes of recombinant protein production (e.g., therapeutic drugs, enzymes). That is because the lactate alters the regulation of biosynthetic enzymes, and can lead to changes in pH in the culture. To mitigate the negative effects of lactic acid accumulation and control the culture pH, chemical ‘base’ is added to the media during the course of a bioprocess. However, the base addition negatively impacts the bioprocess by inhibiting growth and shortening the length of time in which the cells can produce the recombinant protein. This leads to reduced yield, and increased cost-of-goods. Thus, it is of great interest to eliminate lactate production, and UC San Diego researchers have recently developed a new process for achieving this.  

Dual Targeting Agents For Alzheimer's Disease

Alzheimer’s Disease is a prevalent neurodegenerative disorder affecting 10% of people over age 65. It is characterized by a progressive loss of cognitive function and memory impairments that are associated with increased peptide and protein aggregation in the brain. The invention herein describes a novel therapy for Alzheimer’s Disease which would promote the removal of toxic Amyloid-beta peptides out of the brain.

Preventing Protein Aggregation using Thermal Protectants

Protein aggregation in the brain are the causes of the neurodegenerative diseases Alzheimer’s and Parkinson’s. To study diseases and cellular mechanisms, biologists need to be able to efficiently synthesize, isolate and purify proteins. The invention herein is a synthetic nanoparticle (NP) that protects proteins from aggregation at temperatures, which normally cause aggregation. Furthermore, multiple stimuli can release the protein in high yield from the NPs.

A Protein Domain That Protects Ubiquitinated Forms Of Proteins From Degradation In Cis And In Trans

Ubiquitylation affects proteins in many ways, such as activation or inactivation, and signaling for their degradation. It is not fully understood how ubiquitin effects all proteins or how researchers may use it to control cellular processes. This invention describes novel fusion proteins that protect ubiquitylated forms of the target proteins from degradation.

Pyrite Shrink-Wrap Laminate As A Hydroxyl Radical Generator

The invention is a diagnostic technology, as well as a research and development tool. It is a simple, easy to operate, and effective platform for the analysis of pharmaceuticals and biological species. Specifically, this platform generates hydroxyl radicals for oxidative footprinting – a technique commonly employed in protein mapping and analysis. The platform itself is inexpenisve to fabricate, scalable, and requires nothing more than an ordinary pipet to use. In addition, it is highly amenable to scale-up, multiplexing, and automation, and so it holds promise as a high-throughput method for mapping protein structure in support of product development, validation, and regulatory approval in the protein-based therapeutics industry.

A Method For Predicting Glycosylation On Secreted Proteins

Glycosylation is a key post-translational modification that can affect critical properties of proteins produced in biopharmaceutical manufacturing, such as stability, therapeutic efficacy, or immunogenicity. However, unlike a protein's amino acid sequence, glycosylation is hard to engineer since it does not follow any direct equivalent of a genetic code. Despite various attempts to computationally model the process of glycosylation, industrial glycoengineering is still largely carried out using costly and time-consuming trial-and-error strategies and could greatly benefit from computational models that would better meet the requirements for industrial utilization.

MMP-Selective Antibody Inhibitors

Prof. Xin Ge and his colleagues at the University of California, Riverside have developed a library of human, selective MMP-14 antibody inhibitors with nanomolar activities. Unlike known MMP inhibitors, n-TIMP-2 and GM6001, that inhibit a broad spectrum of the MMP family these antibody inhibitors do not exhibit off-target effects with other MMP family members such as MMP-2/MMP-9. Fig. 1 shows inhibition selectivity tests of Fab 3A2, GM6001 and n-TIMP-2. Unlike n-TIMP-2 and GM6001, Fab3A2 shows selectivity for MMP-14 and did not inhibit MMP-2 or MMP-9.     Fig. 2 shows the weight and number of the pulmonary metastic lesions in B16F1 (Mock) and B16F1 MT-MMP (mMT1) mouse models. The mMT1 mouse model treated with the 3A2 (mMT+3A2) antibody showed a reduction in the weight and number of pulmonary metastatic lesions.

Functional Illumination in Living Cells

Researchers at the University of California, Davis, have developed a novel method of developing a wide array of small functional illuminants that do not hinder cell function.

Reversible Chemoenzymatic Protein Labeling

Some of nature’s most complex molecules are made by cellular factories that rely on an acyl carrier protein (ACP) to shuttle growing molecules along biological assembly lines. Post-translational protein modification is important for adding functions to proteins that can be exploited for therapeutics, protein engineering, affinity design and enzyme immobilization, among other applications. Commercial techniques for attaching labels to acyl carrier protein (ACP) and other carrier proteins are currently in use.

A Novel Basic Fibroblast Growth Factor Conjugate for Broad Therapeutic Application

UCLA scientists have developed a novel polymer conjugate of basic fibroblast growth factor (bFGF) for use in commercial and possibly clinical applications. This is the first bFGF conjugate to enhance bioactivity and stability under environmental stressors. 

A Drift-Corrected, High-Resolution Optical Trap

Optical trapping systems are commercially available through several companies. In these systems, the optical trap precision relies on the passive stability of the instrument itself, and therefore demands costly engineering solutions to limit environmental noise that can be coupled into the optomechanical components. Consequently, high-resolution measurements are not possible in common biological laboratory settings that typically lack appropriate vibration isolation and temperature stability.  Researchers at the University of California, Berkeley have developed an invention that addresses a critical problem currently limiting the performance of high-resolution optical traps: that the mechanical drift of optical components often results in physical drift in the location of an optical trap that obscures the displacement-of-interest. The motion of biological motor proteins that are specific to interacting with DNA often take steps along the double helix that is on the order of 0.3 nanometers in size. Accurate measurement of displacements on this scale requires that drift of the trap positions be limited to no more than a few angstroms. However, the current best-performing optical traps suffer from instrumental drift that is almost twice what can be tolerated. Owing to the critical role of these components in all optical trapping systems, and the previously undetectable levels of mechanical drift they undergo, we sought to measure the trap drift with angstrom-level precision using a new approach. This new approach has successfully measured for and corrected for the mechanical drift of these components and demonstrated that this novel invention is capable of consistently reducing the noise floor to levels that have not previously been accomplished.       

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