Showing posts with label Biopharmaceuticals. Show all posts
Showing posts with label Biopharmaceuticals. Show all posts

Monday, April 11, 2016

Biosimilars: They May Look the Same, but One is a Real Stinker


Biosimilars by nature look much like their comparator products, but they sometimes behave quite differently. That’s because the two products often originate from different cell lines or expression systems. And no matter how hard you strive for parity in the way they’re cultured, minor differences in manufacturing can lead to sticky variations in primary amino acid sequence, glycosylation chemical modifications and protein folding issues. The FDA understands biosimilar products sometimes behave differently than their name-brand counterparts. But, they do expect you to understand why those differences occur, and to demonstrate that they have no clinical significance. That’s where ABC Laboratories comes in.

Choosing the Right Analytical Partner May Be the Most Economically Important Decision You Make

FDA guidance calls for a “complete and thorough” CMC section that begins with extensive comparative characterization of the biosimilar and... click here for more.

Tuesday, February 9, 2016

Challenges in the Characterization of Antibody Drug Conjugates


By Glenn Petrie, Ph.D.
Senior Scientific Advisor
ABC Laboratories
www.abclabs.com

Antibody Drug Conjugates (ADC) provide a unique new treatment for a variety of cancers. ADCs consist of a monoclonal antibody (mAb) targeted to the receptor of interest and linked to a highly cytotoxic payload. The mAb binds to the cell and enters the cytoplasm. Once inside the cell, the linker is cleaved and the toxin released. This provides the ability to use highly cytotoxic compounds without the serious side effects of systematic chemotherapy. It is estimated that there are 100-150 ADCs currently in preclinical development. ADCs present unique analytical challenges.

In addition to the complicated mAb, there is the added complexity of combining a cleavable linker and a cytotoxic drug. This introduces the necessity for determining drug loading, linkage sites and Drug Antibody Ratio (DAR). The critical technique for analysis on ADCs is ultra-high resolution QToF mass spectroscopy. This allows for determination of relative loading, DAR and linkage sites, as well as PTMs, disulfide linkages and related substances/degradants (deamidation/oxidation, truncations, and amino acid substitutions). Due to their high resolution, UHR-QTof instruments have the ability to sequence proteins up to 25-30 kDa. The specific protease IdeS (which cleaves just below the hinge region of IgG) under reducing conditions results in three polypeptide chains of ~ 25kDa: Fd region, Fc/2 region and the LC region. Analysis of these digests by UHR QTof MS yields complete sequencing of each polypeptide, DAR, payload and glycan distribution. If necessary, other proteases may be utilized for more detailed analysis. Additional characterization includes the following:

  • Intact mass
  • Deglycosylated mass
  • IEX
  • Imaging CIEF
  • CE-SDS
  • SEC
  • N-linked glycan analysis
  • HIC (secondary DAR analysis)
  • Binding assay
  • Bioassay
  • Higher Order Analysis (CD, AUC, DSC, etc.)

The FDA considers the mAb a drug substance so both the mAb and ADC must be fully characterized. In addition, the agency has been requesting characterization of charge variants of the mAb and ADC. This necessitates preparative IEX followed by characterization of the acidic and basic fractions. Based on all these considerations, characterization of ADCs require careful planning and attention to detail.

Tuesday, January 12, 2016

Well Characterized Biopharmaceuticals Meeting - 2016

By Glenn Petrie, Ph.D.
Senior Scientific Advisor
ABC Laboratories
www.abclabs.com

The 20th annual Well Characterized Biopharmaceuticals (WBCP) meeting will take place in Washington, D.C. the week of January 24th. I attended the inaugural meeting and have only missed one in the last 20 years. This is always an excellent meeting covering both the scientific and regulatory aspects of biopharmaceutical development in equal measure. I am particularly excited about this year’s topics including Higher Order Structure, Combo Products, Biosimilars and Pre-clinical development. I am looking forward to seeing you at the ABC Labs booth to discuss some of our exciting studies. These include:
  • · Utilizing the WES system to quantitate both intact and degraded protein DP in fecal matter
  • · Supported two successful IND submissions for Antibody Drug Conjugates in 2015 with a third in progress. This included full characterization, method development, qualification, release and stability studies. These submissions had NO comments regarding the chemistry section of CMC.
  • · Supported our clients during difficult times
    • During a Phase II trial, reagent changes caused bioanalytical ELISA results to be “offset” from the previous results. We were able to troubleshoot and solve the client’s problem.
    • When a client suddenly severed a relationship with its CRO we were able to transfer, qualify and release methods in support of GMP DS and DP while meeting deadlines.
We hope to speak to you about your biopharmaceutical analytical needs during the breaks, as well as at the Exhibitor Reception, Wednesday, from 5:30-7:00 PM.

Tuesday, September 15, 2015

Gene Editing - A New Frontier


By Glenn Petrie, Ph.D.
Senior Scientific Advisor
ABC Laboratories
www.abclabs.com


A new technology for “editing” genes has scientists excited about an entirely new method for treating disease. The CRISPR technique, along with other gene-editing technologies (TALENS, zinc finger nucleases) allow for the precise deletion of specific DNA sequences. CRISPR involve a Cas9 protein linked to an RNA strand. The RNA sequence serves to target the specific DNA sequence of interest and once bound the Cas9 enzyme cleaves both strands of the DNA.


Above graphic credit (link) to Scitechweb.com 

Since RNA synthesis is relatively cheap and easy this technology has exploded in the last 2 years. Feng Zhang and Eric Lander at MIT have created CRISPR libraries that target essentially the entire human genome. The promise of the technology is to delete genetic mutations and replace them with the correct genes.  Initial targets involve only a single mutation, e.g. sickle-cell anemia, progressing to diseases involving multiple mutations.

As with any new technology there are calls for caution. Chinese scientists reported attempts to repair defective genes in human embryos. This held up the specter of future “designer babies” enhanced in their embryonic stages. Others warn that this technology could be used to “weaponize” bacteria or viruses. Several prominent scientists have proposed a complete moratorium on gene editing until proper safeguards are in place.

Despite these concerns several biotech start-ups have been founded in the last 2 years including Intellia Therapeutics, Editas Medicine and Crispr Therapeutics. Successful in vitro results have been reported for the repair/modification of clinically important genes in hematopoietic stem cells, fibroblasts and T-cells. A quick internet search yields a number of companies offering their services to utilize CRISPR to perform gene editing in both cell lines and whole animals. Based on the extremely rapid advances in this area, gene-editing using CRISPR and other technologies promises to revolutionize our ability to manipulate the genome and attack genetic diseases.

Tuesday, June 23, 2015

A Biotech Scientist Answers the Question: "Is JURASSIC WORLD Possible?"



By Glenn Petrie, Ph.D.
Senior Scientific Advisor
ABC Laboratories
www.abclabs.com


As a boy I was fascinated with dinosaurs. But while most children grew out of this by 7 or 8, I remained engrossed in all things “saurus”. As I started my career in biotechnology, I was particularly interested in Michael Crichton’s book Jurassic Park. I read it in a single sitting and went to the first showing when the movie was released. Now that JURASSIC WORLD is a worldwide phenomenon, interest in reviving extinct species, or de-extinction, has been brought to the forefront. But could JURASSIC WORLD actually be possible? The short answer is no. DNA is the key to any attempts at de-extinction, but while stable, it is not stable for millions of years. Therefore, since dinosaurs went extinct 65 million years ago, there is no viable DNA

However, DNA can survive tens of thousands of years in the right environment, particularly frozen. There are numerous articles in both the scientific and popular literature regarding bringing Wooly Mammoths back to life. Frozen carcasses discovered in Russia have yielded partially intact DNA. JURASSIC WORLD scientists use the bottom-up technique, i.e. start with a fairly complete genome and plug the gaps with that of similar species. In reality the top-down approach is more likely since only partial DNA genomes have been recovered. Scientists are attempting to modify elephant DNA with some of the mammoth’s. The goal is to obtain a hybrid, an elephant with some mammoth traits. With the explosion in biotechnology, one could envision a Pleistocene Park. Rather than dinosaurs it would be populated with animals from the Pleistocene era, ~10,000 years ago. Among the possible exhibits could be Wooly Mammoths, Saber Tooth Cats, Wooly Rhinoceros, Giant Ground Sloths and the Giant Short-faced Bear. Think of the La Brea Tar Pits brought to life.

Even if this is possible, should it be attempted (“But John, if Pirates of the Caribbean breaks down, the pirates don’t eat the tourists.”)? While no one is seriously proposing de-extinction as a theme park, many are interested in using this technology to increase biodiversity and bring back species that humans destroyed, e.g. passenger pigeon, Steller seal, Tasmanian tiger and the dodo. Since there are numerous specimens of these species throughout the museums of the world, there is a reasonable chance of success. However, many ecologists see this as a zero sum game. They believe that any money spent on de-extinction represents less money for efforts to save endangered species; protection, habitat, etc. There may be a middle ground in which de-extinction methods could be used with endangered species to increase genomic diversity and increase the chances for a healthy population.

I look forward to following the latest developments in this exciting field and will provide an update with the release of JURASSIC WORLD 2.

Visit www.abclabs.com for more on Biotech, Biopharmaceuticals and ABC Laboratories' other services.

Thursday, June 11, 2015

ABC Webinar Archive: "Evolving CMC Analytical Techniques for Biopharmaceuticals: Obsolete methods have you feeling on the hook?"


Presented by Glenn Petrie, Ph.D.
Senior Scientific Advisor
ABC Laboratories

Originally produced in association with Contract Pharma
on Thursday, May 28th, 2 pm EST

Regulatory expectations for understanding product impurities and degradants in biopharmaceuticals continue to increase, and many biopharmaceutical developers are finding their “current” CMC methods are quickly becoming obsolete. New, highly sensitive and specific technologies are now emerging from R&D into the QC lab and are becoming the “new normal.” But how do you address heightened regulatory expectations while managing the risk associated with mid-course changes to methods?

To view an archive of the webinar on ABC Laboratories' website, click here.