spacer
spacer
spacer


Or click here to register offline.

Exhibits By:



Media Partners:

       

        

       

Join ISBioTech on:
      
 
PROGRAM CHAIRS:
Alissa M. Resch, PhD - Coriell Institute for Medical Research
Fang Tian, PhD - American Type Culture Collection (ATCC)

Jamie Almeida
Research Biologist, Biochemical Science Division, Bioassay Methods Group
National Institute of Standards & Technology (NIST)
Mouse Cell Line Authentication Consortium Update

Abstract

Misidentified and contaminated cell lines continue to persist in the scientific community. Standard methods are in place for human cell line authentication; however, standards for non-human cell lines are lacking. NIST has partnered with ATCC to validate short tandem repeat (STR) markers developed at NIST for identity testing of mouse cell lines by establishing the Mouse Cell Line Authentication Consortium. This is a collaboration between 13 labs and involves an interlaboratory study to test 50 of the most commonly found mouse cell lines using 19 mouse STR markers. The consortium members were provided a kit containing a master mix, multiplex primers, mouse DNA, a human control, and calibrants. PCR will be used to amplify the target STR markers and these amplicons will be separated using capillary electrophoresis. Each lab will process the samples based on the instrumentation found in their labs which include various models of fragment analyzers, different arrays, and polymers. The fragment analysis data will be analyzed using various software platforms and submitted to NIST upon completion. The results from this consortium will lead to the development of a public STR database for mouse cell lines, and ultimately a written consensus standard for mouse cell line authentication.

Biography

Jamie Almeida received a Bachelor of Science in Microbiology from Arizona State University and a Master of Science in Biotechnology with a concentration in Biodefense from the Johns Hopkins University. She has worked at NIST since 2004 in the Bioassay Methods Group, part of the Biosystems and Biomaterials Division. Some of her past contributions include: 1) stability studies for B. anthracis Sterne spores, 2) development of a rapid bioassay test for ricin using a GFP expressing cell line, and 3) decontamination of various biothreat agents in water systems in the presence and absence of pipe biofilms. Jamie has worked closely with the DNA forensics group at NIST since 2010 learning to separate short tandem repeat alleles using capillary electrophoresis and optimizing multiplex PCR assays. Using this knowledge, she has focused her efforts on the identification of non-human cell lines, specifically monkey, mouse, hamster, and rat.

Joaquina Mascarenhas, PhD
Senior R&D Scientist/Team Lead, MilliporeSigma
Adventitious Agent Risk Mitigation: Engineering
Minute Virus of Mice Resistance into CHO Cells

Abstract

Contamination by the parvovirus minute virus of mice (MVM) remains a continuing challenge in Chinese hamster ovary (CHO) biopharmaceutical production processes. As part of developing a risk mitigation strategy against such events our group has evaluated the genetic engineering of CHO cell lines to create a new host cell line that would be resistant to MVM infection e.g. by inhibiting viral attachment to a cell surface receptor. While the exact functional receptor for MVM binding to CHO cell surface is unknown, previous work in our group has validated the role of sialic acid on the cell surface as important for cell surface binding and internalization of the MVM virus.

Deletion of the CMP-sialic acid transporter Slc35a1 abrogated cell surface binding and internalization of the MVM virus and resulted in complete resistance to MVM infection. Slc35a1 mutants were compared to the wild-type (WT) cells for growth, productivity, and product quality at a 2 L benchtop bioreactor scale. Model recombinant proteins were transfected into the new host cell lines and growth, IgG productivity, and product quality studied. Slc35a1 mutant cell lines were found to be similar to WT cells lines and the monoclonal antibody products, while devoid of terminal sialic acid, had comparable product quality attributes to WT-produced proteins. In more complex glycoproteins, elimination of sialic acid could have a clinical impact. Since MVM preferentially binds to α-2,3 sialylated glycans, we replaced all α-2,3 sialylation with an α-2,6 sialylation phenotype and tested for resistance. Replacing the α-2,3 linked sialic acid with α-2,6 linked sialic acid maintained complete resistance to MVM infection. Gene knockouts targeting viral entry, virus transport, and replication were also tested for MVM resistance. In vitro MVM pull-down assays, whole genome Crispr screens, etc. are ongoing to identify other targets for engineering MVM resistance. As efforts to create better and safer CHO host cell lines continues, the incorporation of viral resistance in host cell lines results in adding greater assurance of production of safely delivered cell-derived products.

Biography

Dr. Joaquina Mascarenhas received her PhD in Biopharmaceutical Sciences from the University of Illinois at Chicago. From there she moved on to a postdoctoral position at Wyeth Biotech (Pfizer) in Andover, Massachusetts. Her current role at MilliporeSigma is as a team lead for research and development focused on next-generation expression systems for the manufacture of recombinant therapeutic proteins and cell-based vaccines. Gene editing tools such as zinc finger nucleases and Crisprs are employed for engineering new host cell lines with superior attributes such as improved product quality, higher productivity, and shorter development timelines. Dr. Mascarenhas is a co-author on publications and is the recipient of patents in the field of CHO host cell engineering related to glycosylation and improved product quality. She is currently leading a team of scientists in the product development of a genetically engineered CHO host cell line that is resistant to viral contamination.

Yvonne A. Reid, PhD
Program Chair Emeritus
Best Practices to Consider for Naming Cell Lines

Abstract

When establishing a new cell line from a tissue sample, or deriving a variant of an existing cell line, it is very important to consider the name or designation of the cell line early in the process. Cell line names that are too long can lead to transposition errors, while cell lines that are too short are not easily searchable and may lead to duplication. Often times a cell line name on the vial in storage may not be traceable to the original tissue or may be different from that used in publications. Changes in the cell line name or inconsistent use of the name oftentimes becomes confusing, leading to mislabeling and misidentification of a cell line and subsequent unreliable data in the scientific field.

Biography

Dr. Reid’s research focuses on the use of DNA hypervariable probes for the intraspecies identification of cell lines. The evolution of this work has led to the implementation of routine screening of all human cell lines by STR analysis. She co-chaired the ATCC Standards Development Organization (SDO) committee on the Development of a Consensus Standard for the Authentication of Human Cell Lines: Standardization of STR Profiling. Dr. Reid has more than 37 years of experience in cell biology, immunology, and molecular biology. As former Head of the Cell Biology Collection, she was responsible for acquisition, expansion, and quality control of new animal cell lines and hybridomas brought into the Cell Biology General Collection. Dr. Reid has authored over 40 peer-reviewed publications and has served on more than a dozen scientific committees, including serving as an ad hoc member of the Comparative Medicine Review Committee. Dr. Reid has served as principal investigator on eight government and non-government contracts. She has been invited to speak, convene, and/or chair sessions at several cell biology conferences sponsored by the American Association for Cancer Research (AACR), ISBioTech, Society for Biomolecular Sciences (SBS), Society for In Vitro Biology (SIVB), International Society for Stem Cell Research (ISSCR), Parenteral Drug Association (PDA), Bioassay and Bioanalytical Methods Development, Select Biosciences-CELLCullture, and Histochemical Society (HCS).

Douglas Storts, PhD
Head of Research, Nucleid Acid Technologies, Promega Corporation
Human Cell Line Authentication
James M. Clinton, PhD
Scientist, ATCC Cell Systems, American Type Culture Collection (ATCC)
Next-Generation Cancer Models: Challenges in Scale-Up
and Cell Banking for Global Distribution
John M. Baust, PhD
President and Lead Scientist, CPSI Biotech
Lisa V. Kalman, PhD
Senior Advisor for Repository Science, Laboratory Research and Evaluation Branch
Centers for Disease Control & Prevention (CDC)
Nahid Turan, PhD
Director, Laboratory Operations, Coriell Institute for Medical Research
Paula Keskula
Operations Leader for the Cell Line Factory, Broad Institute
Fang Tian, PhD
Lead Scientist, Cell Biology Group Leader, ATCC Cell Systems
American Type Culture Collection (ATCC)
Shuo-Hung "Jack" Hsiao, PhD
Scientist, Stem Cell Biology, Coriell Institute for Medical Research


International Society for BioProcess Technology
110 Willow Grove Ct., Norfolk, Virginia 23505 USA • Phone: 757.467.8903, Fax: 757.257.3800
Web: www.isbiotech.org • Email: [email protected]