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Rescooped by H. Fai Poon from TAL effector science
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Microhomology-mediated end-joining-dependent integration of donor DNA in cells and animals using TALENs and CRISPR/Cas9 : Nature Communications : Nature Publishing Group

Microhomology-mediated end-joining-dependent integration of donor DNA in cells and animals using TALENs and CRISPR/Cas9 : Nature Communications : Nature Publishing Group | Biopharm | Scoop.it

(via T. Schreiber, thx)

Nakade et al, 2014

Genome engineering using programmable nucleases enables homologous recombination (HR)-mediated gene knock-in. However, the labour used to construct targeting vectors containing homology arms and difficulties in inducing HR in some cell type and organisms represent technical hurdles for the application of HR-mediated knock-in technology. Here, we introduce an alternative strategy for gene knock-in using transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) mediated by microhomology-mediated end-joining, termed the PITCh (Precise Integration into Target Chromosome) system. TALEN-mediated PITCh, termed TAL-PITCh, enables efficient integration of exogenous donor DNA in human cells and animals, including silkworms and frogs. We further demonstrate that CRISPR/Cas9-mediated PITCh, termed CRIS-PITCh, can be applied in human cells without carrying the plasmid backbone sequence. Thus, our PITCh-ing strategies will be useful for a variety of applications, not only in cultured cells, but also in various organisms, including invertebrates and vertebrates.


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Fesquet didier's curator insight, November 21, 2014 7:55 AM

and other strategy fo Ki experiments

 

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One thing I want every pharma to do... Safety first!

Affymax Biopharm Dissolves After Fatal Drug Reactions. Jun 25, 2014. Affymax Inc., a Cupertino, California-based biopharmaceutical company, has announced that its Board of Directors has approved the liquidation and dissolution of the ...
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Drug maker Dance Biopharm scratches its IPO - Silicon Valley Business Journal

Drug maker Dance Biopharm scratches its IPO - Silicon Valley Business Journal | Biopharm | Scoop.it
Brisbane-based Dance has called off plans for an initial public stock offering, amid investor concerns about potential fallout for U.S. markets from signs of weakness in the global economy.
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Streamlining Antibody Development Using Large Scale, CHO Transient Gene ... - SelectScience.net (press release) (registration)

Streamlining Antibody Development Using Large Scale, CHO Transient Gene ... - SelectScience.net (press release) (registration) | Biopharm | Scoop.it
Search for application articles, scientific posters, methods, techniques and case studies for your laboratory science application on SelectScience.net.
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iBio Grants Caliber License for Production of Antibodies Targeting Ebola - Marketwired (press release)

iBio Grants Caliber License for Production of Antibodies Targeting Ebola - Marketwired (press release) | Biopharm | Scoop.it
NEWARK, DE--(Marketwired - Nov 10, 2014) - iBio, Inc.
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First Chinese-made biologics slated for human trials in the West : Nature Medicine : Nature Publishing Group

H. Fai Poon's insight:

“I was surprised to find that the price of manufacturing biologics in China is about the same as in Europe,” - Chinese company got to find a way to lower their production cost

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Scientists close in on diabetes cure with production of insulin-producing cells - Gizmag

Scientists close in on diabetes cure with production of insulin-producing cells - Gizmag | Biopharm | Scoop.it
Stem cell researchers at Harvard University have created large quantities of human insulin-producing beta cells, which could soon lead to a cure for type 1 ...
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Rescooped by H. Fai Poon from Alzheimer's Disease R&D Review
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Breakthrough Replicates Human Brain Cells for Use in Alzheimer’s Research

Breakthrough Replicates Human Brain Cells for Use in Alzheimer’s Research | Biopharm | Scoop.it
The discovery will allow scientists to quickly, cheaply and easily test drugs that might stop the process in the first place.

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Krishan Maggon 's curator insight, October 13, 2014 2:05 AM
A three-dimensional human neural cell culture model of Alzheimer’s diseaseSe Hoon Choi,Young Hye Kim,Matthias Hebisch,Christopher Sliwinski,Seungkyu Lee,Carla D’Avanzo,Hechao Chen,Basavaraj Hooli,Caroline Asselin,Julien Muffat,Justin B. Klee,Can Zhang,Brian J. Wainger,Michael Peitz,Dora M. Kovacs,Clifford J. Woolf,Steven L. Wagner,Rudolph E. Tanzi& Doo Yeon KimAffiliationsContributionsCorresponding authorsNature (2014) doi:10.1038/nature13800Received 28 January 2014 Accepted 26 August 2014 Published online 12 October 2014
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Finesse Solutions announces launch of SmartVessel for cell culture applications - News-Medical.net

Finesse Solutions announces launch of SmartVessel for cell culture applications - News-Medical.net | Biopharm | Scoop.it
Finesse Solutions, Inc, Santa Clara, CA, a manufacturer of measurement and control solutions for life sciences process applications, announced the launch of beta testing of SmartVessel, a single-use 3 liter bioreactor vessel with integrated...
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Best Stocks on Wall Street: Ibio Inc (NYSEMKT:IBIO), Mead Johnson Nutrition ... - Markets Emerging

Ibio Inc (NYSEMKT:IBIO) skyrocketed 69.84% on Friday, extending gains after it offered assistance to the U.S. government by making its proprietary antibody
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Biosimilars in China: The coming revolution - BioPharma Dive

Biosimilars in China: The coming revolution - BioPharma Dive | Biopharm | Scoop.it
Despite a less rigorous approach to approving biosimilars, China has several approved follow-on biologics—and a growing number of biotechs want to develop biosimilars that meet global standards.
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Eli Lilly confident in its manufacturing for Insulin biosimilar market share - BioPharma-Reporter.com

Eli Lilly confident in its manufacturing for Insulin biosimilar market share - BioPharma-Reporter.com | Biopharm | Scoop.it
Eli Lilly’ manufacturing network will give it the edge in the glargine insulin market, the firm says having received tentative approval of its “Lantus look-alike” from the USFDA.
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New construct for Factor 8!

AbstractBACKGROUND:

Recombinant protein overexpression in mammalian cells constitutes a real challenge in therapeutic protein production. Following the discovery of intron functionality in gene expression, various expression vectors that include them in their sequences have been developed. In this study, the main goal was to develop new lentiviral vectors (LVs) carrying different promoter and intron-containing 5'UTR (5' untranslated region) combinations and the design of LVs for rhFVIII production in Chinese hamster ovary (CHO) cells.

RESULTS:

By combining the human cytomegalovirus (CMV) or the elongation factor 1α (EF-1α) promoters along with different 5'UTRs that included leader introns, between 2 and 12-fold increases were reached, when transient and stable expression of the enhanced green fluorescent protein (EGFP) and rhFVIII were analyzed. Also, new LVs provided with promoters and 5'UTRs from high expression genes, according to a gene database, were designed. Three of them were shown to be superior to the EF-1α promoter in three widely used cell lines.

CONCLUSION:

In the present work, LVs containing different promoters and 5'UTRs were designed. In transient and stable assays some of these constructs have shown higher activity compared with commercial promoters and, therefore, constitute promising candidates for therapeutic protein production.

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Rescooped by H. Fai Poon from Pharma Biotech Industry Review (Krishan Maggon)
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Exploiting the proteomics revolution in biotechnology: from disease and antibody targets to optimizing bioprocess development

Highlights

 

Comparative proteomics identifies novel cancer and disease targets.

Proteomics represent a new tool for mAb discovery.

Proteomics can optimize biotherapeutics development.

Improvements in CHO and E. coli proteome databases aid bioprocessing.

 

Recent advancements in proteomics have enabled the generation of high-quality data sets useful for applications ranging from target and monoclonal antibody (mAB) discovery to bioprocess optimization. Comparative proteomics approaches have recently been used to identify novel disease targets in oncology and other disease conditions. Proteomics has also been applied as a new avenue for mAb discovery. Finally, CHO and Escherichia coli cells represent the dominant production hosts for biopharmaceutical development, yet the physiology of these cells types has yet to be fully established. Proteomics approaches can provide new insights into these cell types, aiding in recombinant protein production, cell growth regulation, and medium formulation. Optimization of sample preparations and protein database developments are enhancing the quantity and accuracy of proteomic results. In these ways, innovations in proteomics are enriching biotechnology and bioprocessing research across a wide spectrum of applications.


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Krishan Maggon 's curator insight, November 3, 2014 7:26 AM
Current Opinion in Biotechnology

Volume 30, December 2014, Pages 80–86

Chemical biotechnology • Pharmaceutical biotechnology

 Exploiting the proteomics revolution in biotechnology: from disease and antibody targets to optimizing bioprocess developmentKelley M Heffner1, Deniz Baycin Hizal2, Amit Kumar1, 3, Joseph Shiloach3, Jie Zhu2, Michael A Bowen2,Michael J Betenbaugh1

 

DOI: 10.1016/j.copbio.2014.06.006

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Which biopharm firms got a head start by winning JumpStart? - Philadelphia Business Journal (blog)

Which biopharm firms got a head start by winning JumpStart? - Philadelphia Business Journal (blog) | Biopharm | Scoop.it
Two suburban Philadelphia life sciences companies will each receive consulting services valued at $50,000.
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UA connection draws biopharm business to town - KGUN 9

UA connection draws biopharm business to town - KGUN 9 | Biopharm | Scoop.it
A Colorado biopharmaceutical company built around an invention developed at the University of Arizona is moving to Tucson.
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The Time Tobacco Actually Saved Lives - Laboratory Equipment

The Time Tobacco Actually Saved Lives - Laboratory Equipment | Biopharm | Scoop.it
The success of an experimental tobacco-based pharmaceutical in saving the lives of two people with Ebola may finally force big pharma to take plant-based drugs seriously.
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Vi-CELL™ XR/Seg-Flow Integrated System for Automated Cell Culture Monitoring - News-Medical.net

Vi-CELL™ XR/Seg-Flow Integrated System for Automated Cell Culture Monitoring - News-Medical.net | Biopharm | Scoop.it
In the bioprocessing industry, automated and real-time cell analysis is still largely unmet and continues to pose a challenge in cell culture monitoring and control.
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New high-performance mammalian expression platform - Phys.Org

New high-performance mammalian expression platform - Phys.Org | Biopharm | Scoop.it
Phys.org Science News Wire : New high-performance mammalian expression platform -- a press release is provided to you ìas isî with little or no review from Phys.org staff.
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Rescooped by H. Fai Poon from Osteoporosis New drugs Review
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Engineering and characterization of a humanized antibody targeting TNF-α and RANKL

Engineering and characterization of a humanized antibody targeting TNF-α and RANKL | Biopharm | Scoop.it

Highlights

 

We engineered and characterized a humanized 8G12 (h8G12) antibody that targeted TNF-α and RANKL.

The h8G12 antibody reduced the TNF-α-mediated apoptosis.

The h8G12 antibody significantly inhibited leukocyte infiltration.

The h8G12 antibody significantly reduced the number of osteoclasts in a dose-dependent manner.

 

Abstract

To neutralize the pathological activities of tumor necrosis factor-α (TNF-α) and receptor activator of NF-κB ligand (RANKL), we engineered and characterized a humanized 8G12 (h8G12) antibody that targeted TNF-α and RANKL. Standard molecular biological and complementarity determining region (CDR)-grafting techniques were used to engineer the h8G12 antibody, and enzyme-linked immunosorbent assays (ELISAs) and Western blotting were employed to determine its binding activation and specificity. TNF-α-mediated cytotoxicity and RANKL-induced osteoclastogenesis assays were used to evaluate the neutralizing effects of the antibody. The cDNA sequences were established by grafting the murine monoclonal antibody (mAb) 8G12 CDRs into the heavy and light chain (HC and LC) variable regions (VH and VL) of the human mAbs 3DGG_B and 1I9R_L, respectively. The recombinant plasmids were transfected into Chinese hamster ovary (CHO) cells to produce the h8G12 antibody, which could simultaneously recognize TNF-α and RANKL. In addition, the h8G12 antibody reduced the TNF-α-mediated apoptosis of L929 cells by 25.84%. Furthermore, the h8G12 antibody significantly inhibited leukocyte infiltration in a murine allergic contact inflammation model. Concurrent with the inhibition of apoptosis, the h8G12 antibody significantly reduced the number of osteoclast-like cells in a dose-dependent manner. These results demonstrated that the h8G12 antibody neutralized the activities of TNF-α and RANKL and that it might be a potential candidate for the treatment of inflammatory bone diseases, such as rheumatoid arthritis (RA).


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Krishan Maggon 's curator insight, October 13, 2014 2:26 AM

Engineering and characterization of a humanized antibody targeting TNF-α and RANKLJun Wanga, b, Yuxuan Dua, Hongyan Qianc, Haitao Yua, Shentao Lia, Xulong Zhanga, Zhanguo Lid,Huihui Yuana, , , Wenming Zhaoa, ,   Biochemical and Biophysical Research Communications

Volume 450, Issue 1, 18 July 2014, Pages 717–722

DOI: 10.1016/j.bbrc.2014.06.046

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My old company is doing all kind of stuff

My old company is doing all kind of stuff | Biopharm | Scoop.it
BioSilta Ltd, based in Cambridge UK, has tied up with Nasdaq-quoted power player Sigma-Aldrich to commercialise crucial technology for scientific researchers.
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World first as Hong Kong surgeons transplant single liver into second patient - South China Morning Post (subscription)

World first as Hong Kong surgeons transplant single liver into second patient - South China Morning Post (subscription) | Biopharm | Scoop.it
World-first procedure sees organ transplanted 11 years ago successfully given to another patient
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Pall ForteBio Introduces Off-The-Shelf Dip And Read Anti-CHO HCP Detection ... - Pharmaceutical Online (press release)

Pall ForteBio LLC, a division of Pall Corp. and a leading supplier of instrumentation and analytics for accelerated drug discovery and development,...
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Enbrel biosimilar biotech Coherus BioSciences sets terms for $85 million IPO - Nasdaq

Enbrel biosimilar biotech Coherus BioSciences sets terms for $85 million IPO - Nasdaq | Biopharm | Scoop.it
Coherus BioSciences, which is developing a biosimilar version of Amgen's Enbrel treatment for inflammatory diseases, announced terms for its IPO on.
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A review on using the right cell line for glycosylation.

A review on using the right cell line for glycosylation. | Biopharm | Scoop.it
H. Fai Poon's insight:
Abstract

Non-human mammalian cells such as CHO have been used predominantly for the production of biopharmaceuticals including monoclonal antibodies (Mabs). Although the glycosylation profile of these products is 'human-like' there is still the possibility of immunogenic epitopes such as α-Gal and Neu5Gc. Human cell lines have now been designed for high productivity of recombinant proteins and ensuring authentic glycosylation patterns. The control of glycosylation on such proteins is important for the efficacy of recombinant biopharmaceuticals as well as the immunogenic properties of viral vaccines such as influenza. We are now starting to understand some of the relationships between the structure of glycans and the function bestowed on the associated protein. This has promoted cell culture technologies for the targeted control of glycosylation to produce pre-determined glycan profiles of secreted products.

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